Novel melon plants

ABSTRACT

The present invention relates to novel plants, in particular to melon plants capable of producing fruits with a new pleasant taste and to seeds thereof. The present invention further relates to fruits of melon plants of the present invention, wherein such fruits have organic acid content, low pH and high sugar contents. The present invention further relates to methods of making and using plants and fruits disclosed herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/535,631, filed Jan. 9, 2004. The above application is incorporatedherein by reference.

FIELD OF THE INVENTION

The present invention relates to novel plants, in particular to melonplants capable of producing fruits with a new pleasant taste. Inparticular, the fruits of the melon plants of the present invention havealtered organic acid contents, lower pH when compared to currentcommercial sweet melon fruits, and high sugar contents.

BACKGROUND OF THE INVENTION

Melon (Cucumis melo L.) is a commercial crop grown worldwide. Cucumismelo is a member of the family Cucurbitaceae. The Cucurbitaceaecomprises about 90 genera and 700 to 760 species, mostly of the tropics.The family includes pumpkins, squashes, gourds, watermelon, loofah andseveral weeds. Cucumis melo L. includes a very wide variety of cultivarsproducing fruits of different shape, external appearance and fleshcolor. Commercial melons generally produce sweet fruits known forexample as Charentais, cantaloupe, honeydew, amarello, Piel de sapo,Kirkagak, Hamy, Ananas, Galia, Oriental that are usually consumed asdessert fruits. Cucumis melo L. also includes non-sweet, commercialcultivars consumed in the Middle to Far East in salad, cooking orpickling, as for example Alficoz, Faqqous, Chito, Conomon (Pitrat et al(2000) Eucarpia meeting Proceedings: 29-36). The taste and aroma ofmelon fruits is determined by a number of factors, including sugars,aroma volatiles, free amino acids, organic acids, pH and solubleminerals (Wang et al. (1996) J. Agric. Food Chem. 44: 210-216). Amongthe four primary tastes (sweet, sour, bitter, salty), sweetness isconsidered to be a very important component of good tasting melonfruits. In commercial melon fruits the sweet taste mostly results fromhigh levels of sucrose (Burger et al. (2002) J. Amer. Soc. Hort. Sci.127(6): 938-943). Sucrose is accumulated at the end of fruitdevelopment, during ripening process (Shaffer et al (1987)Phytochemistry 26: 1883-1887). Melon fruits also initially accumulatehexoses, mainly fructose and glucose, which are the dominant reducingsugars (Stepanski et al (1999) Genetic Resources and Crop Evolution 46:53-62). An important taste component in melon fruits is sweetness, whichis mainly the result of sugars accumulation. Sweetness correlates notonly with the total sugars content, but also with the type of sugars.For example, 1 gram of glucose is the sweet equivalent of 0.7 gram ofsucrose; 1 gram of fructose is the sweet equivalent of 1.7 gram ofsucrose; 1 gram of inverted sugar, i.e. glucose plus fructose generatedfrom 1 gram of sucrose, is the sweet equivalent of 1.3 gram of sucrose(J. A. BABOR et J.IBARZ (1935) Quimica General Moderna).

The flesh of sweet melon fruits has a pH usually above 6.0, but melonaccessions are also known to have a much lower pH, as low as below 5.0.This low pH is widespread over many different melon types as for exampleFaqqous, Chito, Conomon, Momordica, Agrestis (Stepanski et al). In mostof these cases, these melon types combine low pH and low sugar content,e.g. sucrose (Stepanski et al). These fruits are generally not edible infresh consumption without dressing or cooking and, in some cases, theyare even bitter. In most of these melon accessions with low pH, themesocarp, which is the edible part of the fruit, represents a minor partof the total fruit, while the seed cavity and placenta represent a majorpart of the total fruit fresh weight. This is in contrast to sweetdessert melons, where the mesocarp represents a major part of the fruit.Also, in many cases, the fruit size or weight of the melon having low pHis below commercially acceptable ranges.

The fruit flesh of some melons has a sour taste (Kubicki (1962) GeneticaPolonica 3:265-274). The cause for the sour taste remains unclear, butit has been linked with low pH in the fruit flesh (U.S. Pat. No.5,476,998 and Danin-Poleg et al. (2002) Euphytica 125: 373-384). Singlegenes for sour taste (So) and pH have also been reported, although theirgenetic association is not clear (Danin-Poleg et al.).

Attempts have been made to produce melon fruits combining sour and sweettastes. For example, Najd melons based on Arabic wild varieties havebeen reported (Ibrahim and Al-Zeir (1992) HortScience 27: 276-277). U.S.Pat. No. 5,476,998 also described melons with a sour taste, with a meanpH value of 4.8 and a total soluble content of about 11. The melons inU.S. Pat. No. 5,476,998 are derived from MR-1, also known as PI124111(Thomas, Eucarpia '92 pp 142), which is a C. melo Var. Momordica(Roxburg). Fruits of this melon showed low pH (Danin-Poleg et al.). Thismelon variety also involves poor fruit traits such as very mealy,non-aromatic and non-sweet flesh, very climacteric behavior and thinskin bursting at ripening. These undesired traits are difficult toremove by breeding and it is therefore expected that the development ofcommercial products from such a variety would be lengthy and difficult.Accordingly, there is an unmet need for melons producing fruits withalternative or improved tastes. In particular, there is an unmet needfor melon fruits having new combinations of organoleptic characteristicsand aroma.

SUMMARY OF THE INVENTION

The instant invention addresses the need for melon fruits withalternative or improved tastes. Accordingly, the instant inventiondiscloses melon plants capable of producing fruits with novelcombinations of organic acid contents and compositions, pH, and sugarscontents and compositions. The invention also discloses methods ofmaking and methods of using plants of the present invention and theirfruits.

The inventors of the instant application have identified that there is awide variability in the contents and composition of organic acids inmelon fruits. In particular, the inventors of the present invention haveidentified that melon plants produce fruits with varying contents ofcitric acid and varying ratios of citric acid to malic acid.Accordingly, the inventors of the present invention have combinedappropriate content of organic acids, low pH and desired sugar contentsin a melon fruit to obtain novel and extremely pleasant tastes.

Accordingly, in one embodiment, the instant invention discloses melonplants capable of producing fruit with low pH and desired combinationsof citric and malic acid contents, while maintaining or increasinglevels of sugars currently observed in sweet melons. In one embodiment,the instant invention discloses melon plants capable of producing fruitwith low pH and desired combinations of citric and malic acid contents,combined with desired combinations of sucrose and hexoses contents. Inone embodiment, the present invention provides melon plants capable ofproducing fruits with elevated contents of citric acid and lower pH,when compared to current commercial melons. In one embodiment, melonplants of the present invention are obtained by introducing a low pHtrait in a melon plant not comprising said trait. In one embodiment,melon plants of the present invention are obtained by introducing a lowpH gene in a melon plant not comprising said gene. In one embodiment,the low pH gene is obtainable from line IND-35, deposited with NCIMBunder Accession number NCIMB 41202. Accordingly, the present inventiondiscloses melon plants comprising a low pH trait, and producing fruitcomprising desired contents and compositions of organic acids, pH andcontents and compositions of sugars. In one embodiment, increased citricacid contents are achieved while maintaining low levels of malic acid inthe fruit. In one embodiment, fruits of melon plant of the instantinvention have high ratios of citric acid to malic acid.

In one embodiment, the melon plants of the instant invention are capableof producing fruit with citric acid content equal or higher than about400 mg per 100 g fresh weight (fwt). In one embodiment, melon plants ofthe instant invention are capable of producing fruit with a pH of about4.2 to about 5.6. In one embodiment, melon plants of the instantinvention are capable of producing fruit with a sugar content equal orhigher than about 5.0 g per 100 g fwt. In one embodiment, melon plantsof the instant invention are capable of producing fruit with a sugarcontent equal or higher than about 7.0 g per 100 g fwt.

In one embodiment, the present invention discloses melon plants capableof producing fruit with citric acid content equal or higher than about400 mg per 100 g fwt, pH of about 4.2 to about 5.6, and sugar contentequal or higher than about 5.0 g per 100 g fwt.

In one embodiment, the present invention discloses a C. melo plantcapable of producing a fruit comprising at maturity:

a) about 400 mg to about 1,200 mg citric acid per 100 g fwt;

b) pH of about 4.2 to about 5.6; and

c) about 5.0 g to about 15.0 g sugar per 100 g fwt.

In one embodiment, the citric acid content of a fruit of a plantaccording to the present invention is about 400 mg to about 1,000 mgcitric acid per 100 g fwt. In one embodiment, melon plants of thepresent invention produce edible fruits, preferably with a round or ovalshape, and preferably weighting over 450 grams. The flesh of the melonsof the present invention is preferably green, yellow, white or orange.

In one embodiment, the instant invention discloses melon plants capableof producing a very sweet juicy fruit with tart-refreshing sour taste,and referred herein to as “Citric+” plants or fruits. Such fruitscomprise high contents of citric acid and low pH, increasing the acidtaste and providing a tart perception, which covers still too flat andpoor flavors of early mature fruits.

In one embodiment, a fruit of a Citric+plant of the present invention atmaturity comprises:

a) about 600 to about 1,200 mg citric acid per 100 g fwt;

b) pH of about 4.2 to about 5.1; and

c) about 5.0 g to about 15.0 g sugar per 100 g fwt.

In one embodiment, the citric acid content of a fruit of a plantaccording to the present invention is about 600 mg to about 1,000 mgcitric acid per 100 g fwt.

In one embodiment, the present invention discloses a melon plant capableof producing a sweet aromatic fruit with mild-fruity sour taste,referred herein as “Citric−” melon plant or fruit. Such fruits compriserelatively high contents of citric acid and mildly low pH. This mild lowpH makes a fruity sour perception, which covers other still too flat andpoor flavors of early mature fruits.

In one embodiment, the fruit of a Citric−plant of the present inventionat maturity comprises:

a) about 400 to about 650 mg citric acid per 100 g fwt;

b) pH of about 4.6 to about 5.6; and

c) about 5.0 g to about 15.0 g sugar per 100 g fwt.

In one embodiment, the present invention discloses a melon plant capableof producing fruit with low pH and a flesh of deep orange color. In oneembodiment, the present invention discloses a C. melo plant capable ofproducing fruit with pH of about 4.5 to about 5.6, wherein said fruithas orange flesh rated 4 or higher.

In one embodiment, characteristics of melon fruits described herein aremeasured on fruits of melon plants grown in open fields or in plastichouses, and harvested at maturity. In one embodiment fruits areharvested from early maturity to late maturity (stages 2-4, as describedherein). In one embodiment, a fruit of the present invention is atmaturity when its sucrose contents is at or over 2 g sucrose per 100 gfwt.

In one embodiment, the present invention discloses a C. melo plantcapable of producing fruit comprising an acid savor of about 1.6 toabout 3.8 and a sugar savor of about 4.3 to about 5.8. In oneembodiment, the present invention discloses a C. melo plant capable ofproducing fruit comprising an acid savor of about 2.5 to about 3.8 and asugar savor of about 4.3 to about 5.6. In one embodiment, the presentinvention discloses a C. melo plant capable of producing fruitcomprising an acid savor of about 1.6 to about 3.0 and a sugar savor ofabout 5.2 to about 5.8. In one embodiment, the acid savor and sugarsavor are determined by an Expert panel, for example as described inExample 12 herein. In one embodiment, such fruit comprises a pH andcitric acid content as described herein. In one embodiment, such fruitcomprises a pH, organic acid contents and compositions and sugarcontents and compositions as described herein.

In one embodiment, the present invention discloses a C. melo plantcomprising a DNA sequence, which co-segregates with a low pH trait. Inone embodiment, the DNA sequence is a template for amplification of aDNA fragment described herein using the primers described herein. In oneembodiment, the present invention discloses such primers and DNAfragments amplified using these primers. In one embodiment, a DNAfragment described herein is amplified from DNA of said plant using theprimers described herein. The DNA fragments are used as molecularmarkers for a low pH trait. In one embodiment, a DNA fragment of about168 bp to about 178 bp is amplified from the DNA of said plant when theprimers capable of identifying the CMAT141 marker are used. In oneembodiment, a DNA fragment of 168 bp, 173 bp, 169 bp, 172 bp or 178 bpis amplified when the primers capable of identifying the CMAT141 markerare used. In one embodiment, a DNA fragment of less than 176 bp isamplified when the primers capable of identifying the CMAT141 marker areused. In one embodiment, a DNA fragment of less than 175 bp is amplifiedwhen the primers capable of identifying the CMAT141 marker are used. Inone embodiment, a DNA fragment of about 218 bp to about 253 bp isamplified when the primers capable of identifying the NE0585 marker areused. In one embodiment, a DNA fragment of 230 bp, 232 bp, 218 bp, 229bp, 234 bp or 239 bp is amplified when the primers capable ofidentifying the NE0585 marker are used. In one embodiment, a DNAfragment of about 121 bp to about 145 bp is amplified when the primerscapable of identifying the NE1746 marker are used. In one embodiment, aDNA fragment of 124 bp, 127 bp, 133 bp, 142 bp or 145 bp is amplifiedwhen the primers capable of identifying the NE1746 marker are used.

In one embodiment, the present invention discloses a C. melo plantcomprising a DNA sequence, which is a template for amplification of aDNA fragment indicative for the presence of a low pH trait in said plant(acid fragment) or for the absence of the low pH trait (basic fragment)in said plant. In one embodiment, the present invention discloses a C.melo plant comprising a DNA sequence, which is a template foramplification of a basic fragment linked to a low pH trait, wherein saidDNA sequence is linked to said low pH trait. In one embodiment, the C.melo plant comprises such a DNA sequence on one side of a low pH gene.In one embodiment, the C. melo plant comprises such a DNA sequence onboth sides of a low pH gene. In one embodiment, a fruit of such a plantcomprise a pH within the ranges disclosed herein. In one embodiment,fruits of such a plant comprise the contents and compositions of sugarsas described herein. In one embodiment, fruits of such a plant comprisethe contents and compositions of organic acids as described herein. Inone embodiment, fruits of such a plant comprise the pH, contents andcompositions of sugars and contents and compositions of organic acids asdescribed herein.

In one embodiment, the characteristics of a fruit according to thepresent invention described herein remain stable after the fruit reachesmaturity. In one embodiment, such characteristics remain stable after afruit reaches maturity when the fruit is kept on the plant. In oneembodiment, such characteristics remain stable when the fruit isharvested and kept in storage after harvest. This allows for a reducedharvest frequency, and to store or ship a fruit of the present inventionwithout loosing its organoleptic characteristics and aroma. In oneembodiment, the pH of a fruit of the instant invention remains stableafter a fruit reaches maturity. In one embodiment, the citric acidcontent of a fruit of the present invention remains stable after a fruitreaches maturity. In one embodiment, the malic acid content of a fruitof the present invention remains stable after a fruit reaches maturity.In one embodiment, the ratio citric acid to malic acid of a fruit of thepresent invention remains stable after a fruit reaches maturity. In oneembodiment, the pH and organic acid contents and composition of a fruitof the present invention remain stable after a fruit reaches maturity.In one embodiment, such characteristics remain within the rangesdescribed herein after a fruit reaches maturity. In one embodiment, thepresent invention discloses a plant capable of producing a fruit, thecharacteristics of which remain stable for at least 2 days when thefruit is kept on the plant, in one embodiment for at least 3 days whenthe fruit is kept on the plant, in one embodiment for at least 4 dayswhen the fruit is kept on the plant. In one embodiment, the presentinvention discloses a plant capable of producing a fruit, thecharacteristics of which remain stable for at least 5 days when kept instorage at 20° C., in one embodiment for at least 7 days when kept instorage at 20° C., in one embodiment for at least 9 days when kept instorage at 20° C. In one embodiment, the present invention discloses aplant capable of producing a fruit, the characteristics of which remainstable for at least 7 days when kept in storage at 8-12° C. followed byat least 2 days at 20° C., in one embodiment for at least 12 days whenkept in storage at 8-12° C. followed by at least 2 days at 20° C., inone embodiment for at least 26 days when kept in storage at 8-12° C.followed by at least 2 days at 20° C. In one embodiment, a plant of theinstant invention is capable of producing a long shelf-life fruit (LSL)or a medium shelf-life fruit (MSL). In one embodiment, a plant of theinstant invention is capable of producing a non-turning fruit or lowturning fruit. In one embodiment, a plant of the instant invention iscapable of producing a non-climacteric fruit or a low climacteric fruit.

The instant invention further discloses seeds of a C. melo plant of thepresent invention, and seeds of the progeny thereof, wherein saidprogeny seed is capable of producing a plant of the present invention.The instant invention further discloses parts of a C. melo plant of thepresent invention, e.g. ovules or pollen, and fruits of a C. melo plantof the present invention. The instant invention further discloses theflesh of a fruit of a C. melo plant of the present invention. Theinstant invention further discloses the juice of a fruit of a C. meloplant of the present invention.

The present invention further discloses the use of the flesh of a fruitaccording to the present invention in a fresh cut product. The presentinvention further discloses the use of the juice of a fruit according tothe present invention in a soft drink.

The present invention further discloses methods of increasing the citricacid content of a plant comprising obtaining a first C. melo plant;crossing said first C. melo plant with a second C. melo plant comprisinga low pH trait, obtaining a progeny C. melo plant, determining the pHand citric acid content of a fruit of said progeny plant, selecting afruit of said progeny C. melo plant which has increased citric acidcontent, when compared to a fruit of said first C. melo plant. In oneembodiment, said progeny C. melo plant has a lower pH, when compared toa fruit of said first C. melo plant. In one embodiment, the methodcomprises detecting a DNA fragment described herein using the primersdescribed herein. The present invention further discloses the use of amelon plant comprising a low pH trait to obtain a melon plant of thepresent invention. In one embodiment, the melon plant comprising a lowpH trait further has the ability of accumulating relevant levels ofsugars, for example sucrose. In one embodiment, the melon plantcomprising a low pH accumulates high levels of citric acid. In oneembodiment, the melon plant comprising a low pH accumulates low levelsof malic acid. In one embodiment, the melon plant comprising a low pHtrait further has the ability of accumulating relevant levels of sugars,for example sucrose and of accumulating high levels of citric acid andof accumulating low levels of malic acid. In one embodiment, the melonplant comprising a low pH is a plant of line IND-35 or a descendentthereof.

The present invention further discloses a method to produce seed of aplant according to the instant invention comprising obtaining a plant ofthe present invention, self-pollinating said plant or crossing saidplant with another melon plant, and harvesting progeny seed. The presentinvention further discloses a method to vegetatively propagate a melonplant according to the present invention. The present invention furtherdiscloses a method for producing a fruit comprising planting a plantaccording to the present invention, growing said plant and harvesting afruit, wherein said fruit comprises the characteristics describedherein. The method further comprises storing said fruit, for example asdescribed herein. The method further comprises shipping said fruit. Inone embodiment, the characteristics of said fruit described hereinremain stable during the storage of said fruit. In one embodiment, thecharacteristics of said fruit described herein remain stable during thestorage of said fruit. In one embodiment, a plant of the presentinvention is an inbred line, a hybrid, a dihaploid, or a vegetativelypropagated clone.

The present invention thus provides melon fruits offering a pleasantacid component but avoiding an astringent taste. This enhances orcomplements melon flavors to their maximum potential. These combinationsof organic acid content and low pH with high sugars provide new rangesand classes of appealing tastes for fresh consumption or the fresh cutor fresh juice industry. Low pH in fruit flesh also prevents bacterialcontamination in fresh cut and juice industrial processes.

DEFINITIONS

Trait: characteristic or phenotype. For example, in the context of thepresent invention a low pH trait confers a low pH, for example fromabout 4.2 to about 5.6, to the flesh of a melon fruit. A trait may beinherited in a dominant or recessive manner, or in a partial orincomplete-dominant manner. A trait may be monogenic or polygenic, ormay also result from the interaction of one or more genes with theenvironment.

Monogenic: determined by a single locus.

Polygenic: determined by more than one locus.

Dominant: results in a complete phenotypic manifestation at heterozygousor homozygous state.

Recessive: manifests itself only when present at homozygous state.

Partial or incomplete-dominance: when present at the heterozygous stagedetermines a phenotype that is intermediate to that of the homozygousstage or when the trait is absent.

Backcrossing: backcrossing is a process in which a hybrid progeny isrepeatedly crossed back to one of the parents.

Locus: region on a chromosome, which comprises a gene contributing to atrait.

Genetic linkage: association of characters in inheritance due tolocation of genes in proximity on the same chromosome. Measured bypercent recombination between loci (centi-Morgan, cM).

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Isogenic: plants, which are genetically identical, except that they maydiffer by the presence or absence of a gene, a locus conferring a traitor heterologous DNA sequence.

Marker assisted selection: refers to the process of selecting a desiredtrait or desired traits in a plant or plants by detecting one or morenucleic acids from the plant, where the nucleic acid is associated withthe desired trait.

Dihaploid: doubling of haploid (single chromosome) status of the genome(e.g. through anther culture or microspore culture) giving a completehomozygous plant.

“Tester” plant: plant used to characterize genetically a trait in aplant to be tested. Typically, the plant to be tested is crossed with a“tester” plant and the segregation ratio of the trait in the progeny ofthe cross is scored.

Gene: Unit of inheritance. Genes are located at fixed loci inchromosomes and can exist in a series of alternative forms calledalleles.

Allele: One of a pair or series of forms of a gene, which arealternative in inheritance because they are situated at the same locusin homologous chromosomes.

Homozygous: Having like alleles at one or more corresponding loci onhomologous chromosomes.

Heterozygous: Having unlike alleles at one or more corresponding loci onhomologous chromosomes.

Low pH gene: gene, which when present in the genome of a plant leads toa lower pH of the flesh of a fruit of said plant, when-compared to aplant not comprising said gene.

Low pH melon plant: melon plant comprising a low pH trait. In oneembodiment, the pH of a fruit of a low pH melon plant is from about 4.2to about 5.6.

Cucumis melo L: also referred herein to as C. melo or melon.

Cavity: refers to the center of the melon fruit containing seeds andmaternal tissues.

Soluble Solids: refers to the percent of solid material found in thefruit tissue, the vast majority of which is sugars.

Climacteric/non-climacteric: as for example defined in Watkins (2002)“Ethylene synthesis, mode of action, consequences and control” In:Michael Knee (ed) “Fruit Quality and its Biological Basis”. SheffieldAcademic Press, Sheffield, UK. Chapter 8 pp. 180-224, in particular atpage 181, section 8.2.1, first two paragraphs.

Turning melon: turning refers to the marked change in rind color of amelon fruit when it reaches maturity, for example from green to yellowrind in Galia types, or from gray to creamy-yellow rind in Charentaistypes. This change of color is related to the degradation of greenpigments occurring when the fruit reaches maturity.

Non-turning/low-turning melon: the rind color of a fruit of anon-turning or low turning melon does not dramatically change uponmaturation. In the rind of the fruit of non-turning or low turningmelon, there may be a continuous slight increase in yellow component inthe stable background of the rind, as for example in Piel de Sapo typeor in honeydew types. In low-turning melons, a slight pigmentdegradation may also be observed in connection with senescence of thefruit rather than with reaching maturity, which occurs earlier.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides C. melo plants capable of producingfruits with novel tastes. In particular, the present invention providesmelon plants capable of producing fruits with novel combinations oforganic acid contents, pH and sugar contents. The inventors of theinstant application have identified that melon fruits contain variouslevels of organic acid, e.g. citric acid. The inventors of the instantapplication also have identified that melon fruits contain variousrelative contents citric acid and malic acid. Moreover, the inventors ofthe instant application have determined that by introducing a low pHtrait into a sweet melon background, a reduction in pH and an increasein the citric acid content in the fruit are obtained, further expandingthe ability to manipulate the taste of a melon fruit. In one embodiment,melon fruits of the present invention have low contents of malic acid.Accordingly, the present invention discloses melon plants producingfruits having elevated citric acid contents and lower pH, as describedherein. In one embodiment, sugar levels observed in fruits of plants ofthe instant invention are maintained at levels present in currentlyavailable sweet melons or increased. Accordingly, plants of the presentinvention are capable of producing fruits with novel, pleasant tastes.Measurements of pH, citric acid and malic acid contents, and sugarcontents described herein were carried out are shown in Examples 1 to 5.Tables 1A and 1B herein disclose representative melon plants accordingto the present invention and their fruits.

In one embodiment, the melon plants of the instant invention are capableof producing fruit with citric acid content equal or higher than about400 mg per 100 g fresh weight (fwt). In one embodiment, melon plants ofthe instant invention are capable of producing fruit with a pH of about4.2 to about 5.6. In one embodiment, melon plants of the instantinvention are capable of producing fruit with a sugar content equal orhigher that about 5.0 g per 100 g fwt. In one embodiment, the presentinvention discloses melon plants capable of producing fruit with citricacid content equal or higher than about 400 mg per 100 g fwt, pH ofabout 4.2 to about 5.6, and sugar content equal or higher that about 5.0g per 100 g fwt.

In one embodiment, the present invention discloses a C. melo plantcapable of producing a fruit comprising at maturity:

a) about 400 mg to about 1,200 mg citric acid per 100 g fwt;

b) pH of about 4.2 to about 5.6; and

c) about 5.0 g to about 15.0 g sugar per 100 g fwt.

In one embodiment, the fruit comprises about 400 mg to about 1,000 mgcitric acid per 100 g fwt. In one embodiment, the fruit comprises about450 mg to about 950 mg citric acid per 100 g fwt, in one embodimentabout 475 mg to about 900 mg citric acid per 100 g fwt. In oneembodiment, the fruit has a pH of about 4.3 to about 5.4, in oneembodiment about 4.4 to about 5.1. In one embodiment, the fruitcomprises about 5.5 g to about 13.0 g sugar per 100 g fwt. In oneembodiment, the fruit comprises about 7.0 g to about 15.0 g sugar per100 g fwt.

In one embodiment, the ratio citric acid to malic acid in a fruit ofsaid plant is greater than 4.4, in one embodiment greater than 5, in oneembodiment greater than 10. In one embodiment, the ratio citric acid tomalic acid in a fruit of said plant is less than 450, in one embodimentless than 200, in one embodiment less than 150. In one embodiment, afruit of said plant comprises less than about 85 mg malic acid per 100 gfwt, in one embodiment less than about 75 mg malic acid per 100 g fwt.

In one embodiment, the ratio sucrose to hexoses in a fruit of said plantis about 1:1, in one embodiment between about 1:1 and about 1:2, in oneembodiment between about 1:1 and about 2:1.

In one embodiment, the flesh of a fruit of said plant is orange, white,green or yellow. In one embodiment, said fruit is edible in freshconsumption. In one embodiment, the mesocarp of a fruit of said plantrepresents more than 50% of the total fresh fruit weight.

In one embodiment, the C. melo plant comprises a low pH trait. In oneembodiment, the low pH trait is obtainable from a plant of line IND-35,representative seeds of which is deposited under Accession number NCIMB41202, or a descendent of said line IND-35. In one embodiment, the lowpH trait is homozygous or heterozygous in said plant.

In one embodiment of the instant invention, melon plants referred to as“Citric+” plants are disclosed. Such plants are capable of producing avery sweet juicy fruit with tart-refreshing sour taste. Such fruitscomprise high contents of citric acid and low pH, increasing the acidtaste and providing a tart perception. This is compensated by a highsugar content, in one embodiment by elevated levels of reducing sugars(glucose and fructose). The high reducing sugar content is particularlypronounced in early mature stages. The combination of tart and sweettastes, which cannot be obtained by currently available sweet melon, isappreciated by consumers. The tart-refreshing sour sweet meloncombination satisfies this need. This fruity tart perception also coverstill too flat and poor flavors of early mature fruits. In oneembodiment, Citric+plants are generally obtained by introducing a low pHtrait, for example from line IND-35, into an oriental melon background.In one embodiment, the oriental melon background is selected for one ormore of the following criteria: high citric acid content, low malic acidcontent, high citric acid to malic acid content, high sugar content,high hexose content, high ratio hexoses to sucrose, high juiciness.During the introduction of the low pH trait, careful selection for theabove characteristics is maintained until a desired progeny is obtained.

For example, Examples 9, 10 and 11 describe the construction of suchplants. Alternative C. melo plants can also be screened for desiredcharacteristics and are used as starting materials as described herein.

In one embodiment, a fruit of a Citric+plant of the present invention atmaturity comprises:

a) about 600 to about 1,200 mg citric acid per 100 g fwt;

b) pH of about 4.2 to about 5.1; and

c) about 5.0 g to about 15.0 g sugar per 100 g fwt.

In one embodiment, the fruit comprises about 600 to about 1,000 mgcitric acid per 100 g fwt. In one embodiment, the fruit comprises about650 to about 950 mg citric acid per 100 g fwt.

In one embodiment, the pH of the fruit is about 4.4 to about 5.0.

In one embodiment, said fruit comprises about 7.0 g to about 13.0 gsugar per 100 g fwt. In one embodiment, the ratio citric acid to malicacid in a fruit of said plant is greater than 6, in one embodimentgreater than 7, in one embodiment greater than 10. In one embodiment,the ratio citric acid to malic acid in a fruit of said plant is lessthan 450, in one embodiment less than 200. In one embodiment, a fruit ofsaid plant comprising less than about 85 mg malic acid per 100 g fwt, inone embodiment less than about 75 mg malic acid per 100 g fwt, in oneembodiment less than about 60 mg malic acid per 100 g fwt.

In one embodiment, a fruit of a plant of the present invention has agreen or white flesh and at maturity comprises:

a) about 600 to about 1,200 mg citric acid per 100 g fwt;

b) pH of about 4.2 to about 5.1; and

c) about 5.0 g to about 15.0 g sugar per 100 g fwt.

In one embodiment, the fruit comprises about 600 to about 1,000 mgcitric acid per 100 g fwt. In one embodiment, said fruit with green orwhite flesh comprises at maturity about 8.0 g to about 12.0 g sugar per100 g fwt. In one embodiment, said fruit with green or white flesh hasat maturity a ratio citric acid to malic acid between about 25 and 200.In one embodiment the malic acid content of said fruit is less thanabout 50 mg malic acid per 100 g fwt, in one embodiment less than about30 mg malic acid per 100 g fwt. In one embodiment, the ratio sucrose tohexoses in a fruit of said plant is about 1:1, in one embodiment betweenabout 1:1 and about 2:1.

In one embodiment, a fruit of a plant of the present invention hasorange flesh and comprises at maturity:

a) about 600 mg to about 750 mg citric acid per 100 g fwt;

b) pH of about 4.5 to about 5.1; and

c) about 6.0 g to about 13.0 g sugar per 100 g fwt.

In one embodiment, the ratio citric acid to malic acid in said fruit isbetween about 4.4 and 30. In one embodiment, the fruit comprises about7.0 g to about 13.0 g sugar per 100 g fwt.

In one embodiment, the present invention discloses a melon plant capableof producing a sweet aromatic fruit with mild-fruity sour taste,referred herein as “Citric−” melon plant or fruit. Such fruits compriserelatively high contents of citric acid and mildly low pH. This mild lowpH makes a fruity sour perception that cover other still too flat andpoor flavors of early mature fruits and without affecting the fullexpression of fruit flesh flavors and colors of further fully maturefruits. Further during ripening (early to late mature), fruits followrelevant increases of pH, decrease of acidity, in line with the weakbuffer potential from the low citric acid content. This provides fullymature fruits with a very slight sour note to the full expression ofvery sweet and aromatic melon flavors. Melon flavor is based primarilyon sweetness and aromas that usually get the full expression on reallyadvanced fruit ripening stages. Early ripe melons are often described asunpleasant tasteless as sugars and aromas are below expectation and notany other component of flavor is present. In other fruits, such asstrawberry, peach, or oranges, acid taste makes a well appreciatedcomplement in this early ripe stages. The mild-fruity sour and sweetmelon combination avoids or limits the tasteless risk on early ripemelons.

In one embodiment, Citric−plants are obtained by introducing a low pHtrait, for example from line IND-35, into a Charentais melon background.In one embodiment, the Charentais melon background is selected for oneor more of the following criteria: low citric acid content, low malicacid content, relatively high citric acid to malic acid content, highsugar content, high sucrose content. During the introduction of the lowpH trait, careful selection for the above characteristics is maintaineduntil a desired progeny is obtained.

For example, Example 11 describes the preparation of such plants.Alternative C. melo plants can also be screened for desiredcharacteristics and can be used as starting materials as describedherein.

In one embodiment, the fruit of a Citric−plant of the present inventionat maturity comprises:

a) about 400 to about 650 mg citric acid per 100 g fwt;

b) pH of about 4.6 to about 5.6; and

c) about 5.0 g to about 13.0 g sugar per 100 g fwt.

In one embodiment, the fruit comprises about 450 to about 600 mg citricacid per 100 g fwt. In one embodiment, the fruit comprises a pH of about4.8 to about 5.4, in one embodiment to about 5.2. In one embodiment, thefruit comprises about 5.0 g to about 13.0 g sugar per 100 g fwt, in oneembodiment about 6.0 g to about 12.0 g sugar per 100 g fwt. In oneembodiment, the fruit comprises about 7.0 g to about 13.0 g sugar per100 g fwt.

In one embodiment, a fruit of a plant of the present invention hasorange flesh and comprises at maturity:

a) about 400 mg to about 550 mg citric acid per 100 g fwt;

b) pH of about 4.8 to about 5.6; and

c) about 5.0 g to about 11.0 g sugar per 100 g fwt.

In one embodiment, the ratio citric acid to malic acid in said fruit isbetween about 4.4 and 10. In one embodiment, the fruit comprises about7.0 g to about 11.0 g sugar per 100 g fwt. TABLE 1A Summary of data forlines Table 1A discloses representative lines of the present invention.The data shown in Table 1A are averages based on measurements fromindividual trials. The data for the individual trials are disclosed inTables 9-13 in the Examples below. tot. sugar sucrose hexose hex/succitric malic LINES (averages) avg avg avg inv pH avg citric/ avg Plastichouse data nr. frt g/100 g g/100 g g/100 g activity avg mg/100 g malicmg/100 g Green YUSOL High pH 34 9.85 4.50 5.34 1.1 6.05 265 24 11 FleshYUSAZ A Low pH, Citric− 246 8.92 3.24 5.68 1.8 4.67 577 19 31 YUSAZ BLow pH, Citric+ 30 8.70 2.16 6.54 3.0 4.59 832 16 53 White SOLAZ/1 Line71 6.97 2.16 4.81 2.2 4.59 579 23 25 Flesh SOLAZ/2 Line 43 6.19 1.924.27 2.2 4.64 590 47 13 Orange L53AZ A Low pH, Citric− 76 7.09 2.51 4.421.8 4.89 509 3 153 Flesh L53AZ B Low pH, Citric+ 52 8.11 4.24 3.87 0.94.81 701 10 67 L53 High pH 10 8.29 5.05 3.23 0.6 6.67 92 1 93

TABLE 1B Summary of data for hybrids Table 1B discloses representativehybrids of the present invention. The data shown in Table 1B areaverages based on measurements from individual trials.The data for theindividual trials are disclosed in Tables 9-13 in the Examples below.Mehari means the female parent of hybrid MEHARI. tot. sugar sucrosehexose hex/suc citric malic HYBRIDS (average) nr. avg avg avg inv pH avgcitric/ avg Plastic house (PS) & Open Field (OP) data frt g/100 g g/100g g/100 g activity avg mg/100 g malic mg/100 g Green YUSAZ A X YUSOL LowpH, Citric + 17 9.70 4.46 5.24 1.2 4.77 692 46 15 PS FleshMILENIUM-DENEV High pH 12 7.61 3.47 4.13 1.4 5.88 444 X 0 OP F1 WhiteSOLAZ/1 X YUSOL Solaz1 x Yusol 26 9.71 6.40 3.31 0.5 4.75 760 58 13 OPFlesh SOLAZ/2 X YUSOL Solaz2 x Yusol 13 9.08 6.26 2.82 0.5 4.70 782 1626 OP SOLAR F1 High pH 8 8.93 4.39 4.54 1.2 6.48 305 35 9 PS OrangeMehari/L53AZ A Low pH, Citric − 13 7.34 2.24 5.10 2.3 4.98 487 6 78 PSFlesh Mehari/L53AZ B Low pH, Citric + 17 8.35 3.37 4.97 1.5 4.76 663 1160 OP Mehari/L53 High pH 14 8.80 3.96 4.84 1.2 6.31 274 5 54 PS

The instant invention demonstrates that the manipulation of the degreeof sourness in a fruit produce valuable taste variations. Sourness is aresult of the interaction between several parameters, pH being one ofthe most important ones along with organic acids contents andcompositions. The pH greatly influences the degree of dissociation ofacids involved in taste. Each acid has a different dissociation constant(pKa), which refers to the pH, at which 50% of the acid is dissociatedin its relative ions and H+. A lower pH results in more of the acid thatis not in dissociated form. The sourness perception comes mainly fromthe undissociated form of the acid. This explains why, at higher pHlevels, weaker acids such as organic acids with a higher pKa areperceived as sourer than stronger acids.

The pH of a solution is correlated with the concentration and pKa of theacid. Equinormal solutions of stronger acids are sourer than weakerones, since they have a much lower pH. The higher the concentration ofthe acid (titratable acidity), the more sour the acid will be perceived.Sourness is also correlated with the chemical structure of theacidulant, the number of carboxylic groups, the molecular weight, andpolarity of the molecule (see e.g. PURAC Biochem, Gorinchen, TheNetherlands, “Flavor Special”, www.Purac.com). Besides sourness, eachfood acid has its own flavor characteristics in terms of lasting time,flavor intensity and the contribution to other non-sour flavor notessuch as astringency, bitterness and sweetness.

The major organic acids in melon fruits are succinic acid, malic acidand citric acid (Wang et al. (1996) J. Agric. Food Chem. 44: 210-216).Malic acid tends to be associated either with very unripe phases or withsenescence and degenerative phases of fruit over-ripening, and itsdominance in the taste is generally not preferred. In the fruit flesh,gradients of increasing malic acid contents are observed from close tothe seed cavity to close to the fruit rind. Citric acid has a lower pKathan malic acid. Citric acid has more tart and a clean effect, whichoften overpowers other taste or aromas notes. In contrast to malic acid,fruit flesh gradients of decreasing citric acid contents are observedfrom more ripe areas close to seed cavity to areas close to the fruitrind. Such a gradient is also observed for sugars.

Thus, in melon fruits of the instant invention, higher sugar contentsand high citric acid contents tend to be associated in the fruit fleshto provide high contents of both in the most flavorful areas of thefruit, resulting in a very pleasant new taste.

Also, in the melon fruits of the instant invention, the sour perceptionis higher when the pH is low and the citric acid content high.

In one embodiment, fruits according to the instant invention were testedby an Expert panel of trained tasters. Sensory characteristics of thefruits were measured on a scale of quotation from 0 to 9. The pH,organic acid contents and sugar contents of the fruits were alsomeasured. The results of the Expert panel are described in Example 12,Tables 14 and 15. The sensory analysis shows that the acid savor infruits of the instant invention is a function of the pH and of thecitric acid content in the fruit. Based on data gathered from mature andunripe fruits, the Expert panel determined the following formula for thecorrelation between the acid savor and pH and citric acid contents in afruit: Acid savor=13, 12−(2.97×pH)+(0,00587×citric acid content (mg/100g fwt)), with r2=0.70.

Accordingly, in one embodiment, the present invention discloses a C.melo plant capable of producing fruit comprising an acid savor aboveabout 0.5 as determined by an Expert panel. In one embodiment, thepresent invention discloses a C. melo plant capable of producing fruitcomprising an acid savor of about 1.6 or above. In one embodiment, thepresent invention discloses a C. melo plant capable of producing fruitcomprising an acid savor above about 0.5 and a sugar savor of about 4.3or above. In one embodiment, the present invention discloses a C. meloplant capable of producing fruit comprising an acid savor of about 1.6to about 3.8 and a sugar savor of about 4.3 to about 5.8.

In one embodiment, the present invention discloses a C. melo plantcapable of producing fruit comprising an acid savor of about 2.5 toabout 3.8 and a sugar savor of about 4.3 to about 5.6. Hybrids YUSOL XSOLAZ and Mehari X L53 AZ B are representative examples of such plant.

In one embodiment, such fruit comprises a pH and citric acid content asdescribed herein. In one embodiment, the present invention discloses aC. melo plant capable of producing fruit comprising an acid savor ofabout 1.6 to about 3.0 and a sugar savor of about 5.2 to about 5.8.Hybrids YUSAZ X YUSOL, Mehari X L53 AZ A and TD X L53 AZ A arerepresentative examples of such plant.

In one embodiment, such fruit comprises a pH and citric acid content asdescribed herein. In one embodiment, such fruit comprises a pH, organicacid contents and compositions and sugar contents and compositions asdescribed herein.

It is understood that values of Expert panels vary depending on the pH,citric acid content and sugar content of a fruit.

The inventors of the present invention have screened melon entries andaccessions for the content in citric acid and sugars and compositionsand have determined variations within C. melo types (see Table 2). Forexample, some Charentais-type melons were found to have low citric acidcontents (for example Lunastar in Table 2). Some Charentais type fruitswere also found to have a relatively low citric acid to malic acid ratioand a high ratio of sucrose to hexoses. On the other hand, Oriental-typemelons, such as Japanese rocky types melon, were found to have highercontents in citric acid (for example YUCA in Table 2). A representativeof YUCA has been deposited with NCIMB, Aberdeen, AB24 3RY, Scotland onDec. 17, 2003 as YUC-15 under Accession number NCIMB 41203. These melonswere also found to have high ratios of citric acid to malic acid andhigh ratios of hexose to sucrose. Some Galia type melons, such as MG755, were also found to have high citric acid contents. Surprisingly,according to the present invention, in a cross between a Japanese-typemelon and a Charentais-type melon (Prince PF in Table 2), the highcitric acid content of the Japanese-type melon could be disssociatedfrom the high hexose to sucrose ratio and combined with the high sucroseto hexose ratio of the Charentais-type melon. The invention furtherrecognized that variations in citric acid contents also exist amongnon-sweet accessions such as Faqqous and IND-35 (low citric acid contentin Faqqous when compared to IND-35, table 2).

Also, although it was generally assumed in the art that low pH and highsugar accumulation were not frequent or even compatible in the samemelon fruit (Stepanski et al (1999)), the inventors of the presentinvention have been able to combine that characteristics of pH andorganic acid described above with desired sugars contents in the fruits(see e.g. Table 3). TABLE 2 Variation in citric acid contents in variousmelon types and varieties (Fruit analysis data trial Sarrians August1997) No. Citric acid Sucrose Hexoses Glucose Fructose Total Sugar Namefruits Ri pH (mg/100 g) (g/100 g) Glucose (g/100 g) (g/100 g) (g/100 g)Lunastar (Charentais) 6 11.8 6.8 159 6.7 3.2 1.6 1.6 9.2 Yuca-18-3-8 810.5 6.7 286 3.8 4.2 2.3 1.9 8.0 MG 755 OA 4 6.8 294 11.4 3.7 2.1 1.715.1 Prince PF (Japanese X 5 6.1 438 10.0 3.3 1.2 2.1 13.3 Charentais)Sancho 4 409 3.7 4.1 2.1 1.9 7.8 Fagouss Egypt 10 3.8 5.6 35 0.1 1.9 0.91.0 1.9 Fagouss Jim MC Greigt 6 3.3 5.2 62 0.1 2.1 0.9 1.2 2.2 FagoussJordany 11 3.7 5.1 125 0.1 2.1 0.9 1.2 2.2 IND35-1 8 4.5 4.9 340 0.6 3.41.8 1.7 4.0Ri: Brix, refractometer index, measuring soluble solids

TABLE 3 Sugar accumulation vs. pH Statistical analysis of sugaraccumulation (brix) versus pH in F2 populations of a cross betweenIND-35 and YUCA or OGEL (Agadir'98). The analysis shows no negativecorrelation between brix values and the pH. pH < 5.5 pH > 5.5 ANOVARI(Brix) pH RI(Brix) pH P = 0.05 IND35/YUCA-15 NR plts 68 68 22 22 P =0.98 F2 Maximum 13.00 5.40 13.60 7.00 No Significant Average 8.03 4.538.01 5.85 Difference Std. Deviation 2.34 0.30 2.72 0.37 in brix Minimum2.40 3.80 2.40 5.50 Populations IND35/OGEL-17 NR plts 98 98 39 39 P =0.65 F2 Maximum 13.00 5.45 12.20 7.50 No Significant Average 7.75 4.707.93 5.97 Difference Std. Deviation 2.16 0.33 1.89 0.35 in brix Minimum2.80 3.90 4.20 5.50 Populations IND.35 NR plts 29 29 Maximum 7.60 5.40Average 5.45 4.63 Std. Deviation 1.18 0.31 Minimum 3.00 4.10

The present invention further discloses the incorporation of a low pHtrait in sweet C. melo backgrounds. The presence of the low pH trait insweet C. melo backgrounds allowed to lower the pH and increaseconcentrations of organic acids in the fruit flesh and to combinedesirable pH and citric acid contents with appropriate sugarconcentrations and contents, resulting in pleasant, new tastes.

In one embodiment, a low pH trait is determined by a low pH gene. In oneembodiment, a low pH gene is obtained from a wild melon accession orcultivar. In one embodiment, a wild melon accession or cultivar used asdonor for the low pH gene comprises traits that facilitate theconstruction of commercial melons with acceptable agronomiccharacteristics and producing fruits with desirable taste. In oneembodiment, such donor has the ability to accumulate relevant levels ofsugars, such as sucrose. In one embodiment, such wild melon accession orcultivar has at least one of the following characteristics: nonclimacteric behavior, relative bigger fruit size and mesocarp component,crispy flesh.

In one embodiment, the low pH trait is obtained from line IND-35,representative seeds of which was deposited with NCIMB, Aberdeen, AB243RY, Scotland on Dec. 17, 2003 under Accession Number NCIMB 41202. Thisis a C. melo accession from India, which could be classified withinbotanical Var. chito, but it may be better classified as var. acidulus(Naudin, Pitrat et al.) because of its bigger size.

Citric acid is the main organic acid in the fruits of line IND-35 (up to911 mg citric acid per 100 g fwt), while malic acid contents are below50 mg per 100 g fwt. Surprisingly, fruits of IND-35 also have theability to accumulate relevant levels of sucrose (up to 1.7 g sucroseper 100 g fwt and 5.6 g total sugars 100 g fwt after long fruit cycles,i.e after late harvest 50 or 53 days after fruit set). Table 2 alsoreports analysis of fruits of IND-35 showing a pH of about 4.9, a citricacid content of about 340 mg/100 g fresh weight (fwt), and a sugarcontent of about 4.0 g/100 g fwt.

In one embodiment, the low pH trait co-segregates with a molecularmarker. In one embodiment, a molecular marker is a DNA fragmentamplified by PCR, e.g. a SSR marker or a RAPDS marker. In oneembodiment, the presence or absence of an amplified DNA fragment isindicative of the presence or absence of the trait itself or of aparticular allele of the trait. In one embodiment, a difference in thelength of an amplified DNA fragment is indicative of the presence of aparticular allele of a trait, and thus enables to distinguish betweendifferent alleles of a trait. In one embodiment, the instant inventiondiscloses markers, which distinguish between different sources of low pHtrait, and for the presence or absence of a low pH trait in a plant. Forexample, such marker is CMAT 141, described in Danin-Poleg et al. (2001)Theor. Appl. Genet. 102: 61-72 and Danin-Poleg et al. (2002) Euphytica125: 373-384. Other examples of molecular markers are NE0585 and NE1746disclosed herein (see Example 13). These markers are closely linked tothe locus of the pH gene. Markers CMAT141 and NE0585 are on one side ofthe low pH gene, while marker NE1746 is on the other side of the low pHgene.

In one embodiment, the inventors of the instant invention havedetermined that various sources of low pH trait amplify DNA fragments ofdifferent length when the primers for the markers are used (acidfragments, see Example 14, Table 16). For example, in plants of Faggousas fragment of about 176 bp is amplified using for CMAT141. In plants ofaccessions PI414723, PI414724, PI161375 and PI124112 a fragment of about175 bp is amplified. In contrast, in plants of IND-35, described herein,unique fragments of about 168 bp and of about 173 bp are amplified.Using marker NE0585, unique fragments of about 230 bp and about 232 bpare amplified for IND-35. Using marker NE1746, a unique fragments ofabout 127 bp is amplified for IND-35. Another fragment of about 124 bpis amplified for IND-35, which is for example absent in Faggous.

In one embodiment, plants not comprising the low pH trait were alsoanalyzed using the primers described herein. A number of DNA fragmentsassociated with the absence of the low pH trait were determined (basicfragments). For example, using the primers of CMAT141, fragments ofabout 169 bp, about 172 bp and about 178 bp long were detected in plantsnot comprising a low pH trait. Using the primers of NE0585, fragments ofabout 218 bp, about 229 bp, about 234 bp and about 239 bp long weredetected. Using the primers of NE1746, fragments of about 133 bp, about142 bp and about 145 bp long were detected. The skilled person wouldknow how to analyze further plants not comprising the low pH trait anddetermine additional DNA fragments associated with the absence of thelow pH trait. The indicated sizes (in bp) are not absolute but relativeto the other size products detected with the same primer pair. The real(exact) size of the amplified fragments (e.g. determined by sequencing)could be slightly different (+/−1 bp) of those indicated herein.

Accordingly, in one embodiment, the present invention discloses a C.melo plant comprising a DNA sequence, which is a template foramplification of a DNA fragment described herein using the primersdescribed herein. In one embodiment, a DNA fragment of about 168 bp toabout 178 bp is amplified from the DNA of said plant when the primerscapable of identifying the CMAT141 marker are used. In one embodiment, aDNA fragment of 168 bp, 173 bp, 169 bp, 172 bp or 178 bp is amplifiedwhen the primers capable of identifying the CMAT141 marker are used. Inone embodiment, a DNA fragment of less than 176 bp is amplified when theprimers capable of identifying the CMAT141 marker are used. In oneembodiment, a DNA fragment of less than 175 bp is amplified when theprimers capable of identifying the CMAT141 marker are used. In oneembodiment, a DNA fragment of about 218 bp to about 253 bp is amplifiedwhen the primers capable of identifying the NE0585 marker are used. Inone embodiment, a DNA fragment of 230 bp, 232 bp, 218 bp, 229 bp, 234 bpor 239 bp is amplified when the primers capable of identifying theNE0585 marker are used. In one embodiment, a DNA fragment of about 121bp to about 145 bp is amplified when the primers capable of identifyingthe NE 1746 marker are used. In one embodiment, a DNA fragment of 124bp, 127 bp, 133 bp, 142 bp or 145 bp is amplified when the primerscapable of identifying the NE1746 marker are used.

In one embodiment, the inventors of the instant invention have separatedthe link between a DNA fragment indicative of a low pH trait and a lowpH gene. In this case, a DNA sequence, which is a template foramplification of a basic fragment, is linked to a low pH trait.Accordingly, in one embodiment, the instant invention discloses a plantcomprising a low pH trait co-segregating with markers indicative for abasic allele, in particular when using the markers disclosed herein. Inone embodiment, a marker on one side of the pH gene is basic. In oneembodiment, markers on both sides of the pH gene are basic. In oneembodiment, fruits of such a plant comprise a pH within the rangesdisclosed herein. In one embodiment, fruits of such a plant comprise thecontents and compositions of sugars as described herein. In oneembodiment, fruits of such a plant comprise the contents andcompositions of organic acids as described herein. In one embodiment,fruits of such a plant comprise the pH, contents and compositions ofsugars and contents and compositions of organic acids as describedherein.

Accordingly, in one embodiment, the present invention discloses a C.melo plant comprising a DNA sequence, which is a template foramplification of a basic DNA fragment, linked to a low pH gene. In oneembodiment, the present invention discloses a C. melo plant comprising achromosome fragment comprising a DNA sequence, which is a template foramplification of a basic DNA fragment and a low pH gene. In oneembodiment, such chromosome fragment is heterozygous or homozygous insaid plant. Line SOLAZ/2 described herein is a representative example ofsuch a C. melo plant. In SOLAZ/2, fragments of about 172 bp, about 229bp and about 124 bp are amplified using the primers of markers CMAT141,NE0585 and NE1746, respectively.

In one embodiment, the present invention discloses a C. melo plantcomprising a DNA sequence, which is a template for amplification of abasic DNA fragment, on one side of a low pH gene, a low pH gene, and aDNA sequence, which is a template for amplification of a basic DNAfragment, on the other side of the low pH gene, wherein both DNAsequence are linked to the pH gene in said plant. In one embodiment, thepresent invention discloses a C. melo plant comprising a chromosomefragment comprising a DNA sequence, which is a template foramplification of a basic DNA fragment, on one side of a low pH gene, thelow pH gene and a DNA sequence, which is a template for amplification ofa basic DNA fragment, on the other side of the low pH gene. In oneembodiment, such chromosome fragment is heterozygous or homozygous insaid plant. Line SOLAZ/1 described herein is a representative example ofsuch a C. melo plant. In SOLAZ/1, fragments of about 172 bp, about 239bp and about 142 bp are amplified using the primers of markers CMAT141,NE0585 and NE1746, respectively. Line YUSOL/3 described herein isanother representative example of such a C. melo plant. In YUSOL/3,fragments of about 172 bp, about 239 bp and about 145 bp are amplifiedusing the primers of markers CMAT141, NE0585 and NE1746, respectively.

Accordingly, the present invention discloses the use of a melon plantcomprising a low pH trait to obtain a melon plant of the presentinvention. In one embodiment, the melon plant comprising a low pH traitfurther has the ability of accumulating relevant levels of sugars, forexample sucrose. In one embodiment, the melon plant comprising a low pHaccumulates high levels of citric acid. In one embodiment, the melonplant comprising a low pH accumulates low levels of malic acid. In oneembodiment, the melon plant comprising a low pH is a plant of lineIND-35 or a descendent thereof.

In one embodiment, the low pH trait is obtained from a descendent ofsaid line IND-35. The skilled person recognizes that a low pH trait canalso be obtained from other sources. In one embodiment, the low pH traitin such other sources is allelic to the low pH trait in IND-35. In oneembodiment, a test is conducted to determine whether a line to be testedfor a low pH gene comprises an allelic gene to that of line IND-35. LineIND-35 is used as a tester line in a cross with a line to be tested andthe segregation ratio of the low pH phenotype is determined in theresulting progeny.

In one embodiment, a plant of line IND-35 is crossed with breeding melonlines, preferably having high citric acid contents and high levels ofsugar. After each cross plants producing fruits having low pH areselected. Selection is also carried out for increased citric acidcontents and high sugar contents. Examples of the introduction of thelow pH trait into elite lines are disclosed in Examples 7-11.

In one embodiment, a molecular marker as disclosed hereinabove is usedto transfer the pH trait in a desired background, in particular inmethod of increasing the citric acid content of a fruit of a melonplants as described herein. In one embodiment, plants for which afragment corresponding to a low pH trait is amplified are selected andfurther used.

In one embodiment, a reduction of about 1 to about 2.5 pH units, in oneembodiment about 1.5 to about 2.0 pH units, is obtained after theintroduction of the low pH trait in a sweet C. melo plant, when comparedto fruits of a melon not comprising the low pH trait, for example whencompared to an isogenic or near-isogenic line not comprising the low pHtrait. In one embodiment, the citric acid content in the flesh of amelon plant is multiplied by a factor of about 1.5 to about 3 uponintroduction of a low pH trait in said melon plant, when compared tofruits of a melon not comprising the low pH trait, for example whencompared to an isogenic or near-isogenic line not comprising the low pHtrait. Mature fruits of the various plants are compared. For example,Table 1A and 1B show comparisons between melon comprising a low pH traitand melons not comprising a low pH trait, for example between YUSAZ XYUSOL and MILENIUM-DENEV F1, SOLAZ X YUSOL and SOLAR F1, Mehari/L53-L53AZ A-L53 AZ B.

Accordingly, melon fruits of the present invention have higherconcentrations of organic acids than comparable currently availablemelons not comprising the low pH trait (titratable acidity). In melonfruits of the present invention, variations in pH are more correlatedwith the composition (pKa of dominant acids) than with totalconcentration of organic acids. An increase in pH is generally observedduring the late ripening process, and is concurrent with sucroseaccumulation. This increase in pH is buffered by the content in organicacids. Low contents in organic acids result in high increases in pH andreduction in sour taste perception. For example, in melons with lowcitric content as some currently available Charentais-type melons, forexample of climacteric turning type, this increase in pH can be over 1.0pH unit (from pH 6.0 to 7.0). By contrast, higher contents in organicacids lead to reduced pH increases during the ripening process, and thusprevent or reduce the decline in sour taste perception, as for examplein less climacteric, less turning melons.

Accordingly, in one embodiment, the present invention discloses melonsfruits with more stable sour tart taste based on stable low pH and highcitric acid content. In one embodiment, the present invention disclosesmelons fruits with an improved stability of the taste after ripening orpost-harvest. The present invention also discloses a method of delayingor reducing the increase in pH in the fruit of a melon plant comprisingincreasing the content of organic acids in said fruit and introducing alow pH trait in a melon plant. In one embodiment, such method comprisesintroducing the low pH trait in a plant with low or non climactericbehaviour. In one embodiment, such method comprises introducing the lowpH trait in a plant capable of producing a low or non turning fruit.

In one embodiment, the present invention also discloses melons fruits,which mature from an initial early ripe mild sour taste to a fruity andfull flavored taste based on a moderate increase of pH (up to 0.5 pHunits) and moderately elevated citric acid contents.

According to the present invention, a low pH trait is introduced inmelon plants producing fruits with various contents and composition ofsugars. In one embodiment, plants from the Japanese-Oriental type, suchas YUCA, are used as a source of sugars. It is generally observed thataccumulation of sugars, particularly sucrose, is a question of fruitcycle time understood as days after pollination, from fruit setting tofruit ripening. In one embodiment, melon plants producing fruits havingan early and relevant sugar accumulation are used in the instantinvention. For example, plants of the Galia type, such as cultivar OGEL,are used as a source of sugars. In one embodiment, melon plantsproducing fruits having potential for a high and fast accumulation ofsucrose independently from the initial levels of hexoses are used in theinstant invention. For example, plants of the Charentais type, such asL53, are used as a source of such a potential for sucrose accumulation.

Examples of the transfer of a low pH trait to elite lines are disclosedin the Examples below. Table 8 below also discloses melon plantsobtained during the transfer to elite lines. However, other types ofmelons or other melons cultivars or varieties of the types mentionedabove are used in the context of the present invention to constructmelon plants according to the instant invention.

In one embodiment, a C. melo plant comprising a low pH trait is crossedto a C. melo plant of the Oriental type or of the Galia-type. Forexample the C. melo plant comprising a low pH gene is line IND-35. Forexample, the C. melo plant of the Oriental type is YUCA, and the C. meloplant of the Galia-type is OGEL, as described in Example 7 below. Theresulting progenies are for example further crossed to C. melo plants ofthe Oriental type or of the Galia-type to obtain desired pH and contentsand compositions of organic acids and sugars. This process is assistedby measurement of pH, organic acids and sugars, as disclosed herein.Alternative C. melo plants of the Oriental type or of the Galia-type canalso be screened for desired characteristics and used as startingmaterials to obtain C. melo plants as described herein.

In one embodiment, a C. melo plant comprising a low pH trait is crossedto a C. melo plant of the Charentais-type, such as L53 as described inExample 11. Alternative C. melo plants of the Charentais-type can alsobe screened for desired characteristics and used as starting materialsto obtain C. melo plants as described herein.

Other types of C. melo plants are also crossed to a C. melo plantcomprising a low pH trait to obtain melo plants as described herein.

In one embodiment, the present invention discloses a plant capable ofproducing a fruit, the characteristics of which remain stable after afruit reaches maturity and is kept on the plant or when the fruit isharvested and kept in sotrage. In one embodiment, the characteristics ofa fruit described herein remain stable during the commercialpost-harvest life of the fruit. This allows storing or shipping a fruitof the present invention for extended periods of time without loosingits organoleptic characteristics and aromas.

In one embodiment, the pH of a fruit of the instant invention remainsstable after a fruit reaches maturity. In one embodiment, the pH of afruit of the instant invention remains within a range of about 4.2 toabout 5.6. after a fruit reaches maturity. In one embodiment, the citricacid content of a fruit of the present invention remains stable after afruit reaches maturity. In one embodiment, the citric acid content of afruit of the present invention remains at or above 400 mg per 100 g fwtafter a fruit reaches maturity. In one embodiment, the malic acidcontent of a fruit of the present invention remains stable after a fruitreaches maturity. In one embodiment, the ratio citric acid to malic acidof a fruit of the present invention remains stable after a fruit reachesmaturity. In one embodiment, the ratio citric acid to malic acid remainsgreater than 4.4 after a fruit reaches maturity. In one embodiment, suchcharacteristics remain with the ranges described herein after a fruitreaches maturity. In one embodiment, these characteristics remain onwithin about 70% to about 130% of the values of the characteristicsmeasured when the fruit reaches maturity, in one embodiment within about80% to about 120% of the values of the characteristics measured when thefruit reaches maturity, in one embodiment within about 90% to about 110%of the values of the characteristics measured when the fruit reachesmaturity. In one embodiment, the present invention discloses a plantcapable of producing a fruit, the characteristics of which remain stableafter a fruit reaches maturity when kept on the plant, or when harvestedand kept in storage In one embodiment, the present invention discloses aplant capable of producing a fruit, the characteristics of which remainstable for at least 2 days when the fruit is kept on the plant, in oneembodiment for at least 3 days when the fruit is kept on the plant, inone embodiment for at least 4 days when the fruit is kept on the plant.In one embodiment, the present invention discloses a plant capable ofproducing a fruit, the characteristics of which remain stable for atleast 5 days when kept in storage at 20° C., in one embodiment for atleast 7 days when kept in storage at 20° C., in one embodiment for atleast 9 days when kept in storage at 20° C. In one embodiment, thepresent invention discloses a plant capable of producing a fruit, thecharacteristics of which remain stable for at least 7 days when kept instorage at 8-12° C. followed by at least 2 days at 20° C., in oneembodiment for at least 12 days when kept in storage at 8-12° C.followed by at least 2 days at 20° C., in one embodiment for at least 26days when kept in storage at 8-12° C. followed by at least 2 days at 20°C. Typically, after its harvest a fruit may remain under fieldconditions for several hours until its is stored in the conditions setforth herein.

Examples of the evolution of the characteristics of a fruit of a plantof the present invention when kept on the plant are shown in Example 15,Table 17. Examples of the evolution of the characteristics of a fruit ofa plant of the present invention after post harvest storage are shown inExample 16, Tables 18 and 19.

In one embodiment, melons can be described as short shelf life (SLS),medium shelf life (MSL) or long shelf life (LSL). Examples of LSL melonsare Milenium, Piel de Sapo, Italo and non turning Charentais LSL.Examples of MSL melons are Galia and turning Charentais MSL. Examples ofshort shelf life (SSL) melons are classical Charentais.

Typically, the life of a fruit of a SSL melon on the plant is about 1 toabout 2 days. This means that fruits have to be harvested about every 1to 2 days to avoid losses. Typically, the life of a fruit of a MSL melonon the plant is about 3 to about 4 days. This means that fruits areharvested about every 3 to 4 days to avoid losses. Typically, the lifeof a fruit of a LSL melon on the plant is more than about 5 daysTypically a SSL melon can be stored for about 4 to about 7 days at 8-12°C. followed by an aditional 2 days at 20° C., or for about 3 to 4 daysat 20° C. Typically a MSL melon can be stored for about 7 to about 12days at 8-12° C. followed by an aditional 2 days at 20° C., or for about5 to 10 days at 20° C. Typically a LSL melon can be stored for more thanabout 12 days at 8-12° C. followed by an aditional 2 days at 20° C., orfor more than about 10 days at 20° C.

In one embodiment, a plant of the instant invention is capable ofproducing a long shelf-life fruit (LSL) or a medium shelf-life fruit(MSL).

The inventors of the present invention have determined that the low pHtrait and high citric accelerates the climacteric rise in a melon fruitafter a fruit reaches maturity, especially in turning and climactericmelons. This leads to degration of the fruit for example shown by arapid increase in pH and malic acid content and a decrease in citricacid content. This is also shown by the appearance of mealy texture andalcoholic degradation. In turning and climacteric melons, the shelf lifeof the fruit is reduced, while this phenomenon is less perceptible innon-turning or/and non-climacteric melons. Accordingly, in oneembodiment, a plant of the present invention is of a non-turning or lowturning melon genotype. In one embodiment, a plant of the presentinvention is capable of producing a non-climacteric or low-climactericfruit.

Accordingly, in one embodiment, the present invention discloses a C.melo plant with low or non-climacteric behaviour comprising a low pHgene. In one embodiment, a fruit of such C. melo plant further comprisesthe characteristic of sugar contents and compositions described herein.In one embodiment, a fruit of such C. melo plant further comprises thecharacteristic of organic acid contents and compositions describedherein. In one embodiment, a fruit of such C. melo plant comprises aratio of citric acid to malic acid as described herein. In oneembodiment, a fruit of such plant further comprises a pH as describedherein.

In one embodiment, the present invention discloses a C. melo plantcapable of producing a low-turning or non-turning fruit, wherein saidplant comprises a low pH gene. In one embodiment, a fruit of such C.melo plant further comprises the characteristic of sugar contents andcompositions described herein. In one embodiment, a fruit of such C.melo plant further comprises the characteristic of organic acid contentsand compositions described herein. In one embodiment, a fruit of such C.melo plant comprises a ratio of citric acid to malic acid as describedherein. In one embodiment, a fruit of such plant further comprises a pHas described herein.

In one embodiment, the inventors of the instant invention haveidentified that a low pH in the flesh of a melon fruit is associatedwith a very poor color intensity of the flesh of fruits with orangeflesh. In particular, in the case of melon plants producing fruit withorange flesh, a slightly orange or pale orange color was observed (seee.g. Table 4). Surprisingly, the inventors of the present invention havebeen able to combine low pH and deep orange color in the fruit of amelon plant.

Table 4 shows that the color intensity of the fruits of line L53, aparent of Syngenta hybrid MEHARI, is deep to very deep orange (averageof 5.17). Plants originating from a back-cross program to introduce thelow pH trait in L53 but not comprising the low pH trait (L53*High pH)showed a slight decrease in both color intensity and pH (4.71 and 6.46).Plants originating from the back-cross program and comprising the low pHtrait showed a further decrease in color intensity and pH. Plants withthe low pH trait in homozygote stage (L53*Low pH) had the lowest ratefor orange flesh color: 4.04. Plants with the low pH trait inheterozygote stage (L53* acid) have the intermediate rate for orangeflesh color: 4.36. This indicates a linkage or phenotypic effect betweenlow pH and pale orange flesh color. Plants were therefore selected forintense orange flesh color in presence of the low pH trait (rated in oneembodiment at 4 or higher in the scale below, in one embodiment at 5 andhigher, in one embodiment at 6).

In one embodiment, the present invention discloses a C. melo plantcapable of producing fruit with pH of about 4.5 to about 5.6, whereinsaid fruit has orange flesh rated 4 or higher. In one embodiment, theorange color of said fruits is rated 5 and higher, in one embodiment 6.In one embodiment, the pH of said fruit is about 4.5 to about 5.4, inone embodiment about 4.8 to about 5.2. In one embodiment, said fruitcomprises about 400 mg to about 900 mg citric acid per 100 g fwt. In oneembodiment, the fruit comprises about 450 mg to about 750 mg citric acidper 100 g fwt. In one embodiment said fruit comprises a sugar contentequal or above about 5 g per 100 g fwt, in one embodiment about 5.0 g toabout 13.0 g sugar per 100 g fwt. In one embodiment, the fruit comprisesabout 6.0 g to about 12.0 g sugar per 100 g fwt. In one embodiment, theratio citric acid to malic acid in said fruit is between about 4 and 30,in one embodiment between about 5 and about 15. In one embodiment, theratio sucrose to hexoses in a fruit of said plant is about 1:1 and about1:2.

A scale of: 1: white, 2: slightly orange, 3: pale orange, 4: orange, 5:deep orange, 6: very deep orange was used for the color rating (seetable 4). In one embodiment, the color of the fruit is assessed using aspectrophotometer, such as a Minolta CM-2500d spectrophotometer. TABLE 4% of flesh colour intensity class per Fruits Pale Orange Deep Orange Nr.2 3 4 5 6 Col avg 5 True L53 83% 17%  5.17 14 L53*High pH 29% 71% 4.7191 L53* acid 0% 7% 55% 34% 4% 4.36 244 L53* low pH 2% 19%  48% 27% 3%4.04 pH average per flesh colour intensity class Fruits Pale Orange DeepOrange Nr. 2 3 4 5 6 pH avg 5 True L53 6.90 6.64 6.85 14 L53*High pH6.28 6.53 6.46 91 L53* acid 4.75 4.81 4.87 4.94 4.83 244 L53* low pH4.53 4.64 4.76 4.73 4.65 4.72“L53* acid”: means acid phenotype excluded proved fix Low pH homozigotes

In one embodiment, characteristics of melon fruits of the presentinvention are measured on fruits harvested at maturity, i.e. mature orripe fruits. The concrete composition of a melon fruit, and thereforeits taste, is affected by the ripening stage, at which it is harvested.Sugars and organic acids accumulation in melon fruits are dynamicprocesses. As the fruit is approaching maturity their accumulationstarts. As the ripening process advances, each of these compoundsfollows a time-specific pattern of accumulation or degradation, which isalso affected by environmental and growth conditions. The person skilledin the art knows how to recognize a mature melon fruit and understandcriteria defining the maturity of a melon fruit.

In one embodiment, one of the following external maturity markers isused for identification of ripening in sweet melon:

-   -   Senescence of the side fruit leaf (the melon fruit sets in        flower axillar to the peduncle insertion of one leaf named side        leaf fruit). The side fruit leaf becomes necrotic.    -   Fruit skin color changes (turning color green to yellow in Galia        type, gray to creamy-yellow in Charentais type, or increase in        yellow component in Piel de Sapo type).    -   Peduncle dehiscence (specially for Shipper melon, less for        Charentais, less for Galia).    -   Decline in fruit shell firmness, particularly in the blossom end        area.

In one embodiment, “maturity” includes the ripening physiologicalprocess between the stages identified as “2” for “Early-Partial mature”and “4” for “Late-Fully mature”. In one embodiment, the Early-Partialmature stage in melons is identified with the initial accumulation ofsucrose contents at or over 2 g per 100 g fwt of sucrose. In oneembodiment, this is associated with the peaking of citric acid andreducing sugars (glucose plus fructose) contents, and a typical ripefruit flesh pigments and texture. In one embodiment, the Late-Fullymature stage in melons is defined with the end of peaking sucrose sugarscontents. It is also defined as before the degradation process starts,such as loss of fruit flesh consistency, fast declines in glucose,citric acid contents or increases in malic acid contents. Accordingly,in one embodiment, maturity of a fruit starts when the sucrose contentin the fruit reaches 2 g per 100 g fwt. In one embodiment, maturitylasts until no more increase in sucrose content is observed.

The measurements disclosed herein are usually averages of measurementsor data taken from a number of fruits. It is understood that, in anysample, individual fruits of a plant or fruits from individual plants donot lie within the ranges described, because of variations generallyobserved while growing melon plants. In one embodiment, thecharacteristics of melon fruits described herein are measured usingfruits grown in the conditions described herein or under similarconditions (e.g. in Example 6, Tables 5-7 below). In one embodiment, afigure for a characteristic according to the instant invention is anaverage taken from fruits grown from plastic house staked plants (onefruit per plant in staked plants).

In one embodiment, a plant of the instant invention is an inbred line, adihaploid or a hybrid. In one embodiment, an inbred line comprises a lowpH trait and the characteristics of organic acids, pH and sugarsdescribed herein. In this case, such inbred line is crossed with anothermelon plant, preferably another inbred line, to obtain a hybrid plantaccording to the instant invention. In one embodiment, the other inbredline in the cross is also capable of producing fruit having high citricacid contents and/or high sugar contents. Representative inbred linesaccording to the present invention are disclosed in Table 1A.

In one embodiment, a plant of the instant invention is a hybrid plant.In this case, some of the characteristics of low pH, high citric acidcontent and high sugar content are contributed from one of the parent,while the remaining ones are contributed by the other parent. In a oneembodiment, one parent in the cross produces fruit having high citricacid content, high sugar content but having high pH (e.g. around pH6.5), while the other parent in the cross produces fruit having low pH.In one embodiment, fruits of the other parent also have high citric acidcontent. Representative hybrids according to the present invention aredisclosed in Table 1B. As used herein, the term “plant” includes plantcells, plant protoplasts, plant cell of tissue culture from which melonplants can be regenerated, plant calli, plant clumps, and plant cellsthat are intact in plants or parts of plants, such as pollen, flowers,leaves, stalks, and the like. In one embodiment, a plant of the instantinvention is capable of producing edible melon fruits. The mesocarprepresents the edible part of the melon fruit (flesh). The mesocarpsurrounds the seed cavity, which is itself surrounded by the rind (orshell). The mesocarp of a fruit according to the present inventionpreferably has a thickness of more than 2 cm and preferably representsmore than 50% of the total fruit fresh weight. In one embodiment, theflesh of a fruit of a melon plant of the present invention has a green,white, yellow or orange flesh.

In one embodiment, the present invention provides regenerable cells foruse in tissue culture of a plant of the present invention. The tissueculture is capable of regenerating plants having the characteristics ofa plant of the present invention. Preferably, the regenerable cells insuch tissue cultures are immature embryos, protoplasts, meristematiccells, callus, pollen, leaves, anthers, roots, root tips, or flowers.Still further, the present invention provides melon plants regeneratedfrom the tissue cultures of the invention. Examples of regenerationprotocols are disclosed in U.S. Pat. No. 6,420,631. The presentinvention further provides a method of asexually propagating a plant ofthe present invention comprising collecting a tissue of a plant of thepresent invention, cultivating said tissue to obtain proliferatedshoots, rooting said proliferated shoots to obtain rooted plantlets. Thepresent invention further discloses a method to produce seed of a plantaccording to the instant invention comprising obtaining a plant of thepresent invention, self-pollinating said plant or crossing said plantwith another melon plant, and harvesting progeny seed. The presentinvention further discloses a method for producing a fruit comprisingplanting a plant according to the present invention, growing said plantand harvesting a fruit, wherein said fruit comprises the characteristicsdescribed herein. The method further comprises storing said fruit, forexample as described herein. The method further comprises shipping saidfruit. In one embodiment, the characteristics of said fruit describedherein remain stable during the storage of said fruit. In oneembodiment, the characteristics of said fruit described herein remainstable during the storage of said fruit.

Some charateristics of a number of C. melo types and lines are describedbelow. These characteristics are examples for the various melon typesand lines, and are not meant to be limiting but are illustrative of thevarious melon types and lines. Variations from these characteristics mayoccur.

Melon Type Charentais:

Fruit: Shape round to hight round; Size 600 to 1200 grs; Rind:White-Gray with Green-Gray sutures, smooth skin (smooth or slightlynetted skin). Orange, melty, very aromatic sweet flesh, climacteric.Some Charentais varieties have reduced climateric phase and not turningrind colour.

Var LUNASTAR: Lunastar is monoecious hybrid from Nunhems in Charentaistype.

Melon Type Japanese:

Fruit: Shape round to hight round; Size 700 to 1500 grs; Rind:White-Gray netted skin, (exceptionally sutured). Green-Yellow(exceptionally orange), from cryspy to melty and few aromatic and verysweet flesh, only in cases. Long cycle non climacteric.

Var YUCA: Andromonoecious hybrid in Japanese type green flesh.

Var PRINCE: Andromonoecious hybrid in Japanese type orange flesh.

Melon Type Galia:

Fruit: Shape round to hight round; Size 600 to 1500 grs; Rind: Greentuning yellow netted skin. Green-White, melty, aromatic sweet flesh.Short cycle Medium shelf life, climacteric. Var MG.755: Andromonoecioushybrid in Galia type. Very short fruit cycle with high brix.

Melon Type Piel De Sapo:

Fruit: Shape Ovoid to oblong-eliptic; Size 2 to 5 Kgrs; Rind: GoldenGreen spotted, few longitudinal netted skin. White, crispy, juicy andaromatic sweet flesh. Medium fruit cycle Long shelf life, Nonclimacteric.

Var Sancho:Andromonoecious hybrid in Piel de sapo type. Golden rind withhigh brix.

Melon type Fagous:

Fruit: Shape Medium short cilindrical; Size 300 to 1000 grs; Rind: Greenskin. White, crispy, non sweet flesh. For comsumption green in salad.Monoecious

IND 35: Fruit: Shape Long pear-ovoid; Size 300 to 1000 grs; Rind: LightGreen spoted skin yellow at maturity. White, crispy, non sweet flesh.For consumption green in salad. Monoecious. Syngenta Seeds line, nonclimacteric.

YUSOL: Shape round to hight round; Size 600 to 1300 grs; Rind: Yellowfew netted with light green sutures skin. Green-white, melty, fewaromatic sweet non acid flesh. Short cycle Medium shelf life, nonturning low climacteric. Andromonoecious Syngenta Seeds Line.

YUSAZ A: Shape round to hight round; Size 600 to 1300 grs; Rind:White-gray few netted with Green-Gray sutures skin. Green, speciallycrispy, few aromatic, sweet and acid flesh. Short cycle Medium-long selflife, non turning, low to medium climacteric. Andromonoecious SyngentaSeeds Line.

YUSAZ B: Shape round to hight round; Size 600 to 1300 grs; Rind: Yellownetted with Green-Gray sutures skin. Green, specially crispy, fewaromatic, sweet and acid flesh. Long cycle, long self life, non turninglow climacteric. Andromonoecious Syngenta Seeds Line.

MILENIUN-DENEV: Shape round to hight round; Size 600 to 1500 grs; Rind:Yellow netted non sutured skin. Green-White, crispy, non aromatic, nonacid and sweet flesh. Long cycle, non turning, long self life, nonclimacteric. Andromonoecious Syngenta Seeds Hybrid.

SOLAZ/1: Shape round to hight round; Size 700 to 1400 grs; Rind: Yellowfew netted with light green sutures skin. White, crispy, few aromaticsweet and acid flesh. Short cycle, long self life, non turning, very lowclimacteric. Andromonoecious Syngenta Seeds Line.

SOLAZ/2: Shape round to hight round; Size 600 to 1300 grs; Rind: Yellowfew netted with light green sutures skin. White, crispy, few aromaticsweet and acid flesh. Short cycle long self life, non turning, very lowclimacteric. Andromonoecious Syngenta Seeds Line.

MEHARI: monoecious hybrid from Syngenta in Charentais type. Turningmedium shelf life.

L53: andromonoecious line from Syngenta in Charentais type. Turningmedium shelf life. Small size round flat shape. Derived from varLUNASTAR, a monoecious hybrid of Nunhems in turning Charentais type.

TD: monoecious line from Syngenta in Charentais type. Non turning andlong shelf life. Derived from var TORNADO, a monoecious hybrid fromLimagrain in non turning Charentais type.

All references cited herein are incorporated by reference in theapplication in their entireties. When ranges are disclosed herein, it isunderstood that all the individual numbers falling within these rangesare also part of the invention.

The following examples are intended to provide illustrations of theapplication of the present invention. The following examples are notintended to completely define or otherwise limit the scope of theinvention.

EXAMPLES Example 1

Preparation of Melon Extracts

A wedge of ca. 400 g was taken from a melon fruit, the seeds and theskin (1 cm thickness) were removed. The flesh was cut in small parts,which were blended for 30 seconds in a Warring blender until a smoothslurry was obtained. The slurry was filtrated over a Whatman paperfilter, the juice centrifugated in an Eppendorf centrifuge at 10.000 gand stored at −20 degrees Celcius.

Example 2

Determination of Citric Acid Content

Samples as prepared in Example 1 were incubated with citrate lyase (CL)to convert citric acid to oxaloactate and acetate. In the presence ofthe enzymes malate dehydrogenase (MDH) and lactate dehydrogenase (LDH),oxaloacetate and its decarboxylated derivative pyruvate were reducedwith NADH to respectively L-malate and L-lactate. The decrease of NADHis proportional with the quantity of citric acid in the sample and canbe determined at 340 nm. The assay was conducted in microplates. 20 μlof diluted sample was added to a microtiterplate. 200 μl of assaymixture containing NADH, MDH and LDH was added and mixed to the plate.The reaction was started with 15 μl of start-solution containing CL. Theplate was mixed and let the reaction proceed for 1 hour. Absorbancevalues were measured at 340 nm with a microtiterplate reader (BiotekEL808 reader with KCJunior software and computer). A calibration curvewas used to calculate the concentration citric acid in the samples. Theenzymes were purchased from Roche Diagnostics.

Example 3

Determination of Malic Acid Content

Samples as prepared in Example 1 were incubated with L-malatedehydrogenase (MDH) and NAD⁺ to convert L-malate to oxaloacetate. Theequilibrium of the reaction is on the side of malate but was forced tothe side of oxaloacetate with hydrazine by means of derivatization. NADHformed during the assay is proportional with the malic acid content inthe samples. NADH can be determined at 340 nm as a measure for malicacid. The assay was conducted in microplates. 20 μl of diluted samplewas added to a microtiterplate. The reaction was started by adding 200μl of assay mixture containing hydrazine, MDH and NAD⁺ at pH 10. Theplate was mixed and the reaction was allowed to proceed for 1 hour.Absorbance values were measured at 340 nm with a microtiterplate reader(Biotek EL808 reader with KCJunior software and computer). A calibrationcurve was used to calculate the concentration malic acid in the samples.MDH was purchased from Roche Diagnostics.

Example 4

Determination of Glucose Fructose and Sucrose Contents

Glucose:

Glucose was determined with the enzymes hexokinase andglucose-6-phosphate dehydrogenase (G-6-PDH). Glucose was phosphorylatedwith hexokinase to glucose-6-phosphate (G-6-P) and subsequentlydehydrated to 6-phosphogluconate with the aid of NADP and G-6-PDH(reactions 1 and 2, respectively). The concentration of formed NADPH(the H-acceptor) is quantitatively related to the initial glucoseconcentration and was measured at 340 nm in the UV range of the lightspectrum.

Fructose:

Fructose was determined in the same assay. Fructose was phosphorylatedto fructose-6-phosphate (F-6-P) with the enzyme hexokinase. F-6-P wasconverted to G-6-P with the enzyme phosphoglucose-isomerase (PGI) andsubsequently in 6-PG as described in reaction 2 above.

Sucrose:

Sucrose was converted to glucose and fructose with the enzymeβ-fructosidase. Formed glucose was determined according to the reactions1 and 2 as described above. The determinations were conducted inmicrotiterplates, absorbance values are measured with a microtiterplatereader (Biotek ELx808 with data collection software and computer).Enzymes were purchased from Roche Diagnostics.

Example 5

Measurements of pH

The pH of samples as described in Example 1 was determined using aCRIMSON GLP21 pHmeter calibrated to pH 4 and pH 7 with standardsolutions.

Example 6

Growth Conditions of Melons Plants

Melon plants were grown under different conditions at differentlocations (see table 5 below). The dates of the trials are shown inTable 6 and the growing conditions in the different trial are describedin Table 7. TABLE 5 Description of trials LOCATION IN SPAIN CROP TYPECONDITIONS TORREPACHECO EL EJIDO VINE BEES STACKED HAND PLASTIC-SUBSTRATE T.St T.St. POLLINATION POLLINATION OPEN FIELD HOUSE SOILHYDROPONIC FA02PS X X X X SP03PV X X X X SP03PS X X X X SP03OF X X X XSU03PS X X X X FA03PS X X X XFA: Fall,SP: Spring,SU: Summer,PS: Plastic house staked crop,PV: Plastic house vine crop

TABLE 6 Trial dates TRIAL DATES SOWING TRANSPLANTING 1ST POLLINATIONMEDIUM HARVEST FA02PS Aug. 1, 2002 Aug. 19, 2002 Sep. 1, 2002 Oct. 30,2003 SP03PV Jan. 18, 2003 Feb. 18, 2003 Apr. 4, 2003 May 15, 2003 SP03PSFeb. 18, 2003 Mar. 18, 2003 Apr. 18, 2003 May 30, 2003 SP03OF Mar. 10,2003 Apr. 11, 2003 May 20, 2003 Jun. 28, 2003 SU03PS Jul. 24, 2003 Aug.8, 2003 Aug. 20, 2003 Oct. 6, 2003 FA03PS Aug. 7, 2003 Aug. 22, 2003Sep. 8, 2003 Nov. 3, 2003Herein, SP03OF1 refers to analytical data of fruits harvested on Jun.20, 2003. SP03OF2 refers to fruits harvested July 4, 2003.

TABLE 7A Growing conditions for FA02PS Location: EL EJIDO T.St.PLASTIC-HOUSE Week no. 2002 29/30/31/32 33/34/35/36 37/38/39/4041/42/43/44 45/46/47/48 temperature-day 31.5 31 28.6 22.4 18.4temperature-night 25 23.1 22.4 17.8 14.4 temperature-24 hours 28.8 27.425.5 19.8 18 R.Humidity-day % 51 47 55 78 84 R.Humidity-night % 73 72 7889 90 R.Humidity-24 h. % 60 69 66 84 88Weeks are numbered starting on January 1, week 29 being the 29^(th) weekof the year.

TABLE 7B Growing conditions for SP03PV, SP03PS, SU03PS, FA03PS Location:EL EJIDO T.St., PLASTIC-HOUSE Week no. 2003 1/2/3/4 5/6/7/8 9/10/11/1213/14/15/16 17/18/19/20 21/22/23/24 25/26/27/28 temperature-day 15.916.7 25.1 26.9 26.9 31.2 37.4 temperature-night 10.4 11.1 17.9 20.3 21.925.6 28.6 temperature-24 hours 12.6 13.5 21.4 23.8 24.8 28.9 33.9R.Humidity-day % 71 59 46 53 63 54 38 R.Humidity-night % 91 83 64 68 7367 57 R.Humidity-24 h. % 83 73 55 61 67 59 45 Week no. 2003 29/30/31/3233/34/35/36 37/38/39/40 41/42/43/44 45/46/47/48 temperature-day 37.130.9 25.3 20.8 19 temperature-night 28.5 25.1 21.8 17.9 15temperature-24 hours 33.5 28.3 23.6 19.2 16.7 R.Humidity-day % 32 60 8289 86 R.Humidity-night % 46 80 89 96 99 R.Humidity-24 h. % 38 69 86 9293

TABLE 1C Growing conditions for SP03OF TORREPACHECO T.St. OPEN FIELDWeek no. 2003 9/10/11/12 13/14/15/16 17/18/19/20 21/22/23/24 25/26/27/28Temperature Max 28.0 36.0 34.0 33.0 37.0 Temperature avg. Max 19.9 26.227.0 28.0 34.5 Temperature Med 14.3 20.1 19.9 22.1 27.1 Temperature avg.Min. 8.8 14.0 12.7 16.2 19.7 Temperature Min. 5.0 8.0 9.0 12.0 17.0

Example 7

Transfer of the Low pH Trait to Elite Lines

1. Line IND-35 was crossed with a selected set of Syngenta elite linesin the El Ejido Trial Station (Spain).

The selected lines were:

a).-YUCA- 15: Selected because of high potential for sugars accumulationin medium long cycle (time from fruit set to fruit ripening). YUCA is aproprietary Syngenta Seeds line obtained through 5 generations ofself-pollination. It is a Japanese Rocky melon type, non turning andvery low climacteric.

b).-OGEL-17: Selected because of medium short cycle (time from fruit setto fruit ripening) and medium high potential for sugars accumulation.This line is a proprietary Syngenta Seeds line obtained through 10generations of self-pollination from a breeding F1 from cross betweenparental lines of commercial hybrids GUSTAL and RADICAL (SyngentaSeeds). It is a GALIA melon type, turning and medium climacteric.

F1 crosses produced self-pollinated F2 descendent generation. The F2fruits were selected for low pH and sugar (Table 8A).

2. Both F2 populations were grown and cross pollinated plant by plantwith a new selected set of Syngenta elite lines:

a).-IND35/YUCA-15 F2 population was crossed with MG.755-68 (755), aproprietary Syngenta Seeds line obtained through 5 generations ofself-pollination from a commercial hybrid BETULO (Syngenta Seeds). It isa GALIA turning and medium climacteric melon type selected for thepurpose because of green flesh, high sugars, short cycle and relativelyhigh citric acid content.

b).-IND35/OGEL-17 F2 population was crossed with SEN19C8 (SN8). Thisline is a proprietary Syngenta Seeds line, non turning and very lowclimacteric, obtained through 6 generations of self-pollination of acommercial hybrid EARLS SEINU (Yae Nogey Seeds Co. Isahaya, Nagasaki,Japan). It is a Japanese Rocky melon type selected for the purposebecause of high sugars and netting.

In these populations, the single dominance genetic regulation for theLow pH trait was verified and that no linkage existed between sugaraccumulation and said Low pH. (Table 3). From each populations (150plants each), recombinants including the higher R.I. Brix degree and thelowest pH were selected. Cross-pollinated seed progeny was collected.

The cross progenies selected from F2 population a) included the plantsidentified as: 755YUCIND-19, 755YUCIND-49, 755YUCIND-75.

The one selected from F2 population b) included the plant identified as:SN8OGLIND-03.

755YUCIND lines were selected for sucrose accumulation. SN8OGLIND wasselected for citric acid accumulation. These cross progenies were grownand produced data for pH, sugars and organic acid contents. Linescombining high sugars, low pH and high citric acid content wereselected. The analysis of the selected plants is shown in Table 8B.TABLE 8 Analysis of intermediates Table 8A: Analytical Data extract forselected plants in F2 (Agadir Autumn 1998) Geno FEMALE pH BRIX typeYUCA-15.071.xIND35-1)-49. 4.30 13.0 Aa YUCA-15.071.xIND35-1)-75. 4.1012.4 Aa YUCA-15.071.xIND35-1)-19. 4.05 10.4 AA OGEL-17.974.xIND35-1)-03.4.57 11.8 Aa

TABLE 8B Analytical Data extract for progeny B1F1 (El Ejido Spring 1999)Citric acid Malic acid Glucose Fructose Sucrose Total sugar Brix Nr.MALE FEMALE pH (mg/100 g) (mg/100 g) (g/100 g) (g/100 g) (g/100 g)(g/100 g) R.I. Fruits SN8 OGLIND-03. 4.7 773 52 2.3 2.4 1.7 6.3 10.0 54.5 930 48 1.8 2.0 4.4 8.2 12.0 4.7 809 56 2.0 2.2 2.4 6.6 10.2 755YUCIND-49. 5.0 540 45 2.6 2.7 1.7 6.9 10.0 5 5.0 619 39 1.7 2.4 3.3 7.411.5 4.8 535 67 2.0 2.5 1.8 6.3 9.9 755 YUCIND-75. 5.0 549 38 2.5 2.72.7 7.9 12.0 3 5.4 564 52 2.6 2.7 2.2 7.4 11.0 5.3 568 40 2.3 2.5 2.67.5 10.7 755 YUCIND-19. 5.5 624 44 1.5 2.2 4.5 8.2 12.0 8 4.7 648 53 1.92.0 3.7 7.6 13.0 5.1 569 44 1.7 2.1 3.3 7.2 11.1

TABLE 8C Analytical Data extract for progeny B2&3F1 (El Ejido Autumn1999) Citric acid Glucose Fructose Sucrose Total Sugar Brix Nr. MALEFEMALE pH (mg/100 g) (g/100 g) (g/100 g) (g/100 g) (g/100 g) R.I. FruitsIOTYU 4.7 802 2.5 2.5 2.9 7.9 12.5 4.7 1036 2.3 2.4 3.7 8.4 14.5 755 5.1594 2.5 2.6 2.4 7.4 12.0 YUCA64 SN8 OGLIND-03. 4.7 771 2.3 2.4 2.4 7.112.6 9 755 YUCIND-19. 755)2 YUCIND-19.

TABLE 8D Analytical Data extract for progeny B3F1&2 (Torrepacheco Spring2000) Citric acid Glucose Fructose Sucrose Total Sugar Brix Nr. MALEFEMALE pH (mg/100 g) (g/100 g) (g/100 g) (g/100 g) (g/100g) R.I. FruitsSOLAR-19 IOTYU SN8 OGLIND-03. SOLAR-19 IOTYU SN8 OGLIND-03. 4.14 725 1.91.6 15.0 13 IOTYU IOTYU SN8 OGLIND-03. 4.73 523 1.8 1.2 14.3 3 YUCA64755)2 YUCIND-19. 4.87 566 2.3 1.7 14.7 3

TABLE 8E Analytical Data extract (El Ejido Autumn 2000) Citric acidGlucose Fructose Sucrose Total Sugar Brix Nr. MALE FEMALE pH (mg/100 g)(g/100 g) (g/100 g) (g/100 g) (g/100 g) R.I. Fruits SOLAZ4.4.4 4.32 5733.0 1.2 7.0 SOLAZ4.7.2. 4.44 564 3.3 1.1 11.0 SOLAZ2.4.2 4.52 494 2.91.4 11.0 SOLAZ2.4.5 4.43 474 2.7 1.0 10.0 YUCA-40 IOTYU)2 SN8 OGLIND-03.4.48 453 2.5 1.0 12.0 YUCA64 755)2 YUCIND-19. 4.79 904 4.8 1.8 13.0

TABLE 8F Analytical Data extract (El Ejido 2001) Citric acid GlucoseFructose Sucrose Total Sugar Brix Nr. MALE FEMALE pH (mg/100 g) (g/100g) (g/100 g) (g/100 g) (g/100_(g)) R.I. Fruits SOLAZ 1 4.59 940 2.3 2.94.1 9.4 11.0 SOLAZ 2 4.63 770 2.5 2.7 1.3 6.6 8.0 YUSOL27 YUCA-40 IOTYSNC8OGLIN 4.6 741 3.4 2.5 4.6 10.4 14.0 U)2 D-03. YUSOL27 YUCA-64 755)2YUCIND-19. 5.05 467 2.5 2.6 5.0 10.1 12.0 L53 755 YUCIND-75. 4.80 7181.3 2.2 1.6 5.1 7.0 L53 755 YUCIND-49. 4.68 673 2.5 2.7 2.3 7.6 9.0

Example 8

Trait Introgresion Into SOLAZ

The Low pH trait was introgressed in white flesh non climacteric LSLmelons with round shape and yellow sutured skin.

1. Selected SN8OGLIND- 103 progeny plants were crossed with IOTYU, aproprietary Syngenta Seeds line obtained through generations ofself-pollination from the traditional open pollinated Japanese varietyMakuwauri EIJYU (Nanto seed Co. Ltd, Kashiwara, Nara, Japan). It wasselected because of non-climacteric, non turning, very high sugarcontent, high citric acid content, yellow rind.

2. Selected plants of the progeny from previous cross were crossed withSOLAR-19 a proprietary Syngenta Seeds line obtained through 5generations of self-pollination from a commercial hybrid SOLARKING F1(Nunhems Zaden BV, Haelen, Holland). It is a long shelf life, nonturning Galia melon type selected for the purpose because of nonclimacteric, very high sugar content, white flesh, yellow and nettedrind.

Seven cycles of self-pollination were carried out from these crossprogenies. The selection and fixation on the self-pollination progenieswas done with analytical data for pH, sugars and organic acid contents(Table 8C and 8D). Fixed lines progenies were selected:

A) SOLAZ 1 selected as Low pH in combination for high sugaraccumulation, powdery mildew tolerance, and plant with Summer growingadaptation (Table 8E and F).

B) SOLAZ 2 selected as Low pH with combination for medium sugaraccumulation, and plant with Spring growing adaptation (Table 8E and F).

3. Progenies SOLAZ 1 and SOLAZ 2 were used as males pollinators forcrosses with three proprietary Syngenta Seeds lines, YUSOL 1, 2 & 3,obtained through 6 generations of self-pollination from a breedinghybrid MD.997 F1 from cross YUCA-15×SOLAR-48 both proprietary SyngentaLines with origin indicated before.

The following F1 combinations were obtained:

A) YUSOL 1/SOLAZ 1; when using SOLAZ 1 as pollinator of lines YUSOL 1.

B) YUSOL 2/SOLAZ 1; when using SOLAZ 1 as pollinator of lines YUSOL 2.

C) YUSOL 3/SOLAZ 2; when using SOLAZ 2 as pollinator of lines YUSOL 3.

The analysis of these selected plants and commercially availablecontrols is shown in Tables 9 and 10. The plants were grown under theconditions described in Example 6. The number of fruits tested in eachexperiment is shown (nr. Frt). The measurements were carried out asdescribed in Example 1-5. The figures for sucrose (suc), hexoses (hex),and total sugars are in g per 100 g fresh weight (fwt). The figures forcitric acid and malic acid are in mg per 100 g fresh weight (fwt). TABLE9A SOLAZ tot. tot. hex/suc Growing nr suc suc hex hex sugar sugar inv pHpH citric citric citric/ malic malic conditions frt avg stdev avg stdevavg stdev activity avg stdev avg stdev malic avg stdev SOLAZ/2 SP03PS 62.41 1.70 3.41 0.34 5.82 1.42 4.6 0.2 605 98 42 14 21 FA03PS 37 1.430.81 5.12 0.36 6.55 1.34 3.58 4.7 0.2 576 86 52 11 15 Average 43 1.924.27 6.19 2.50 4.6 590 47 13 SOLAZ/1 SP03PS 4 2.90 0.86 3.93 0.42 6.831.35 4.6 0.0 549 81 16 33 15 FA03PS 35 1.85 1.18 5.88 0.33 7.73 1.603.17 4.5 0.1 663 107 35 19 17.5 SOLAZ/1 SP03PS 4 2.01 0.22 3.82 0.205.83 1.90 4.6 0.1 513 53 20 26 11 FA03PS 16 1.80 0.74 5.32 0.27 7.131.25 2.95 4.5 0.1 591 83 52 11 10 SOLAZ/1 SP03PS 5 2.35 0.88 3.79 0.376.14 1.61 4.8 0.2 497 58 14 36 24 FA03PS 7 2.02 1.36 6.14 0.35 8.15 1.653.04 4.5 0.1 664 86 28 24 24 Average 71 2.16 4.81 6.97 2.34 4.6 579 2725 average SOLAZ/1 and /2 114 2.10 4.68 6.77 4.6 582 27 22

TABLE 9B YUSOL tot. tot. hex/suc Growing nr suc suc hex hex sugar sugarinv pH pH citric citric citric/ malic malic conditions frt avg stdev avgstdev avg stdev activity avg stdev avg stdev malic avg stdev YUSOL/1SP03PS 8 4.25 1.18 4.77 0.82 9.02 1.12 6.0 0.3 295 85 47 6 10 FA03PS 85.01 0.58 5.33 0.85 10.34 1.16 1.07 6.3 0.1 214 49 13 16 10 Average/1 164.63 5.05 9.68 1.09 6.1 255 30 11 YUSOL/2 SP03PS 6 2.58 0.62 4.99 0.817.57 1.94 5.8 0.1 297 83 424 1 3 FA03PS 1 3.92 7.30 11.21 0.54 5.9 344 X0 Average/2 7 3.25 6.14 9.39 1.24 5.9 320 1 YUSOL/3 SP03PS 6 5.04 1.434.04 0.81 9.08 0.80 6.1 0.2 225 97 12 18 21 FA03PS 5 6.23 0.67 5.64 0.4711.87 0.54 0.91 6.2 0.2 218 62 9 24 10 Average/3 11 5.63 4.84 10.47 0.856.2 221 11 21 Average 1, 2, 3 34 4.50 5.34 9.85 1.06 6.05 265 24 11

TABLE 10A Hybrid SOLAZ X YUSOL tot. hex/suc Growing nr suc suc hex hexsugar inv pH pH citric citric citric/ malic malic conditions frt avgstdev avg stdev avg activity avg stdev avg stdev malic avg stdev SOLAZ/1X SP03PV 4 5.58 1.07 3.38 0.51 8.95 0.61 4.8 0.1 845 87 11240 0 2YUSOL/1 SP03OF1 5 6.25 0.43 3.71 0.70 9.96 0.59 4.7 0.1 794 25 76 10 12SP03OF2 5 6.90 0.60 2.55 0.19 9.44 0.37 4.8 0.1 679 50 47 14 8 average/114 6.24 3.21 9.45 0.52 4.8 773 97 8 SOLAZ/1 X SP03PS 3 3.45 0.24 4.680.52 8.14 1.36 4.7 0.1 697 32 45 16 12 YUSOL/2 SP03OF1 5 9.20 0.34 2.850.19 12.05 0.31 4.7 0.1 800 27 42 19 7 SP03OF2 4 7.01 1.30 2.69 0.409.70 0.38 4.8 0.1 744 46 39 19 4 average/2 12 6.56 3.41 9.97 0.68 4.7747 42 18 SOLAZ/2 X SP03PV 4 4.41 0.72 3.20 0.44 7.61 0.73 4.7 0.2 86362 100 9 18 YUSOL/3 SP03OF1 4 4.94 2.05 3.23 0.29 8.17 0.65 4.5 0.1 78985 277 3 4 SP03OF2 5 9.42 1.09 2.04 0.37 11.46 0.22 4.9 0.1 694 63 109 62 average/3 13 6.26 2.82 9.08 0.53 4.7 782 162 6 Average 39 6.35 3.159.50 0.58 4.7 767 70 11

TABLE 10B Controls tot. tot. hex/suc Growing nr suc suc hex hex sugarsugar inv pH pH citric citric citric/ malic malic conditions frt avgstdev avg stdev avg stdev activity avg stdev avg stdev malic avg stdevSOLAR F1 SP03PV 6 5.28 0.93 2.52 0.32 7.80 0.92 0.48 6.1 0.08 500 51 2818 26 FA03PS 2 3.50 0.05 6.55 0.17 10.06 0.22 1.87 6.9 0.04 109 17 X 0 0Average 8 4.39 4.54 8.93 1.17 6.48 305 35 9 MILENIUM SP03PV 4 2.99 1.234.94 0.67 7.93 1.57 1.65 5.8 0.1 546 25 X 0 0 SP03OF1 5 2.29 0.15 4.160.10 6.44 0.15 1.82 5.8 0.05 441 23 X 0 0 SP03OF2 3 5.15 3.82 3.30 0.998.45 4.81 0.64 6.1 0.4 345 40 X 0 0 Average 12 3.47 4.13 7.61 1.37 5.88444 X 0

Example 9

Trait Introgresion Into YUSAZ A.

The Low pH trait was introgressed in green flesh non climacteric LSLmelons with round shape and yellow sutured skin.

1. Direct back-cross was made from line 755YUCIND-19 using as recurrentparent, line MG.755-68 indicated above.

Selected plants of the progeny from previous cross were crossed withYUCA-64 a proprietary Syngenta Seeds line obtained through 5 generationsof self-pollination from a same origin as the sister line indicatedabove.

This line was selected because of green flesh, non turning, nonclimacteric long-shelf life, very high sugars very crispy watermelontexture and sutures.

Three cycles of self-pollination from this cross progenies were carriedout. The selection and fixation on the self-pollination progenies wasmade with analytical data for pH, sugars and organic acid contents(Table 8C-E).

2. Selected plants from progeny generated above were crossed withYUSOL.3 a proprietary Syngenta Seeds line described above. This line wasselected because of green flesh, shorter cycle, very high sugars, yellowrind and sutures.

Five cycles of self-pollination from the previous cross progenies wasmade. Selection and fixation breeding process concentrated on theself-pollination progenies for short cycle, green flesh, crispywatermelon texture and yellow rind. Selection and fixation was assistedwith analytical data for pH, sugars and organic acid contents. Therewere so selected and fixed lines progeny YUSAZ A.

3. YUSAZ A lines were used as males pollinators for crosses with threeproprietary Syngenta Seeds lines, YUSOL 1 a proprietary Syngenta Seedsline described above. The F1 combination: YUSOL 1/YUSAZ A was obtained.

The analysis of these selected plants is shown in Table 11. The plantswere grown under the conditions described in Example 6. The number offruits tested in each experiment is shown (nr. Frt). The measurementswere carried out as described in Example 1-5. The figures for sucrose(suc), hexoses (hex), and total sugars are in g per 100 g fresh weight(fwt). The figures for citric acid and malic acid are in mg per 100 gfresh weight (fwt). TABLE 11A YUSAZ A tot. tot. hex/suc Growing Nr sucsuc hex hex sugar sugar inv pH pH citric citric citric/ malic malicconditions frt avg stdev avg stdev avg stdev activity avg stdev avgstdev malic avg stdev YUSAZ A/1 SP03PS 16 3.01 1.28 5.36 0.67 8.37 1.784.6 0.2 602 90 28 22 13 FA02PS 3 2.05 0.43 5.62 0.37 7.67 2.74 4.6 0.1655 135 21 31 1 SU03PS 6 3.70 1.00 5.76 0.53 9.46 1.92 1.55 4.6 0.2 514115 23 23 18 FA03PS 43 2.10 0.73 7.01 0.44 9.11 1.25 3.34 4.6 0.2 558 8244 13 18 Average 68 2.72 5.94 8.65 2.35 4.6 582 29 22 YUSAZ A/2 SP03PS25 3.81 0.99 5.03 0.54 8.84 1.32 4.8 0.2 586 55 17 35 14 FA02PS 6 3.530.83 5.74 0.38 9.27 1.63 4.6 0.1 582 70 10 56 15 SU03PS 95 3.99 1.424.82 0.41 8.80 1.72 1.21 4.8 0.3 543 80 16 33 19 FA03PS 52 3.70 1.306.12 0.35 9.81 1.28 1.66 4.7 0.1 578 63 18 33 13 Average 178 3.76 5.439.18 1.46 4.7 572 15 39 average YUSAZ A/1, /2 246 3.24 5.68 8.92 1.904.7 577 19 31

TABLE 11B F1 Hybrids using YUSAZ A tot. tot. hex/suc Growing Nr suc suchex hex sugar sugar inv pH pH citric citric citric/ malic malicconditions frt avg stdev avg stdev avg stdev activity avg stdev avgstdev malic avg stdev F1 Hybrid SP03PS 12 5.24 1.83 5.09 0.99 10.33 0.974.9 0.2 659 113 69 10 7 YUSAZ A1/ YUSOL F1 Hybrid SP03PS 5 3.68 0.465.40 0.63 9.08 1.47 4.6 0.1 724 87 35 21 8 YUSAZ A2/ YUSOL Average 174.46 5.24 9.70 1.22 4.8 692 46 15Characteristics of line YUSAZ A are shown. YUSAZ A/1 and YUSAZ A/2represent two sister lines. The F1 hybrids result from a cross with lineYUSOL.

Example 10

Trait Introgresion Into YUSAZ B.

The Low pH trait was introgressed in green flesh non climacteric LSLmelons with round shape and yellow sutured skin.

1. Selected plants of cross SN8OGLIND-103×IOTYU progeny indicated abovewere back-crossed again with IOTYU, non turning and very lowclimacteric, green flesh melon (see Table 8D).

Selected plants of the progeny from previous cross were crossed withYUCA-40 a proprietary Syngenta Seeds line obtained through 5 generationsof self-pollination from a same origin as the sister line indicatedabove (see Table 8E).

This line was selected because of green flesh, non climactericlong-shelf life, very high sugars very crispy watermelon texture andsutures.

2. Selected plants from progeny generated after one cycle ofself-pollination from the one before were crossed with YUSOL.3 aproprietary Syngenta Seeds line described above. This line was selectedbecause of green flesh, shorter cycle, very high sugars, yellow rind andsutures (see Table 8F).

Five cycles of self-pollination from the previous cross progenies werecarried out. Self-pollination progenies were selected for green flesh,crispy watermelon texture, high citric acid content and yellow rind.

3. The selection and fixation was assisted with analytical data for pH,sugars and organic acid contents. Fixed lines progeny YUSAZ B wereselected (Table 12).

YUSAZ B is an example of a “Citric+” melon plant.

YUSAZ B lines were used as males pollinators for crosses with threeproprietary Syngenta Seeds lines, YUSOL 1 a proprietary Syngenta Seedsline described above. The F1 combination: YUSOL 1/YUSAZ B was obtained(Table 12).

The plants were grown under the conditions described in Example 6. Thenumber of fruits tested in each experiment is shown (nr. Frt). Themeasurements were carried out as described in Example 1-5. The figuresfor sucrose (suc), hexoses (hex), and total sugars are in g per 100 gfresh weight (fwt). The figures for citric acid and malic acid are in mgper 100 g fresh weight (fwt). TABLE 12 YUSAZ B tot. tot. hex/suc Growingnr sucr suc hex hex sugar sugar inv pH pH citric citric citric/ malicmalic conditions frt avg stdev avg stdev avg stdev activity avg stdevavg stdev malic avg stdev YUSAZ B/0 SP03PS 7 3.16 0.66 4.99 0.60 8.141.58 4.7 0.1 891 73 17 73 12 FA02PS 2 2.46 0.01 5.75 0.42 8.20 2.34 4.30.0 848 14 11 75 2 FA03PS 6 1.61 0.61 6.63 0.39 8.24 1.02 4.12 4.4 0.1878 169 11 78 57 Average 15 2.41 5.79 8.19 2.68 4.5 872 13 75 YUSAZ B/1SP03PS 6 1.81 1.20 6.74 0.40 8.55 3.71 4.7 0.2 696 133 39 18 17 FA02PS 12.72 6.76 9.48 2.48 4.7 739 18 41 FA03PS 8 1.21 0.45 8.39 0.20 9.60 0.696.94 4.7 0.1 939 37 26 36 16 Average 15 1.91 7.30 9.21 4.38 4.7 791 2832 average YUSAZ B/0, /1 30 2.16 6.54 8.70 3.53 4.6 832 16 53Characteristics of line YUSAZ B are shown. YUSAZ B/1 and YUSAZ B/2represent two sister lines. The F1 hybrids result from a cross with lineYUSOL.

Example 11

Trait Introgresion to MEHARI F1

The low pH trait was introgressed in orange flesh melons. The selectedtarget was the conversion of the commercial hybrid MEHARI (SyngentaSeeds) into two low pH versions, one with higher citric acid content andone with lower citric acid content.

1. Direct backcrosses from lines 755YUCIND-49 and 755YUCIND-75 wereconducted using as recurrent parent line L53, a proprietary SyngentaSeeds. L53 is a Charentais melon type selected for the purpose becauseof high sugars, low citric content. L53 is a parent of the commercialhybrid MEHARI (see Table 8F).

During the backcross process and within the progeny of each cross plantswith low pH and more or less acid perception (higher or lower citriccontent) were selected, in addition to other traits to be closer to therecurrent L53.

After the second back-cross (third cross) the following progenies wereobtained:

a) L53)3)×755YUCIND75-15/03/06/, selected for low pH and low citric acidcontent

b) L53)3+)×755YUCIND49-8/2/03/05/, selected for low pH and high citricacid content. Four cycles of self-pollination from these cross progeniesand one further back-cross with L53 were carried out. The selection andfixation breeding process on the self-pollination progenies was assistedwith analytical data for pH, sugars and organic acid contents.

2. Fixed lines progenies were selected:

A) L53AZ A selected from previous population a) and fixed for low citricacid content and low pH and, for other traits, close to the recurrentL53.

Line L53AZ A is an example of a “Citric−” melon plant.

B) L53AZ B selected from previous population b) and fixed for low pH andhigh citric acid content and, for other traits, close to the recurrentL53.

Plants were also selected for intense orange flesh color in presence ofthe low pH trait. The intensity of the orange color in the flesh had tobe particularly selected for, as a generally poor orange color intensitytended to be associated within low pH acid in the fruit flesh (see Table4).

Progenies lines L53AZ A and L53AZ B, and line L53 were used in crosseswith the other parent line of MEHARI to obtain MEHARI AZ A, MEHARI AZ Band MEHARI the current commercial F1, respectively.

All these products, lines and crosses were tested in agronomic trialsand analytical data were collected from fruits (Table 13).

The plants were grown under the conditions described in Example 6. Thenumber of fruits tested in each experiment is shown (nr. Frt). Themeasurements were carried out as described in Example 1-5. The figuresfor sucrose (suc), hexoses (hex), and total sugars are in g per 100 gfresh weight (fwt). The figures for citric acid and malic acid are in mgper 100 g fresh weight (fwt). TABLE 13A MEHARI, low pH/high pH tot. tot.Hex/suc Growing nr suc Suc Hex hex sugar sugar inv pH pH citric citriccitric/ Malic malic conditions frt avg Stdev Avg stdev avg stdevactivity avg stdev avg stdev malic avg stdev Male high pH FA02PS 4 4.111.66 2.93 0.67 7.04 0.71 6.9 0.2 65 26 1 114 38 cit/− (L53) SP03PS 31.98 0.85 2.70 2.05 4.68 1.36 6.2 0 52 11 1 55 26 FA03PS 6 9.07 0.674.07 0.40 13.14 1.06 0.45 6.9 0.2 159 40 1 109 18 Average/High pH− 105.05 3.23 8.29 6.7 92 1 93 Male acid cit/− SP03PS 4 2.75 1.65 3.85 0.386.61 1.40 5.0 0.3 442 168 4 109 51 (L53AZ A) SU03PS 24 2.03 1.40 3.351.11 5.82 1.83 1.65 4.8 0.3 484 132 3 150 64 FA03PS 48 2.76 1.85 6.070.62 8.82 2.19 2.20 4.8 0.4 600 158 3 200 130 Average/low pH, citric− 762.51 4.42 7.09 4.9 509 3 153 Male acid SP03PS 3 4.82 1.37 2.72 0.30 7.540.56 5.0 0.1 654 86 9 75 23 (L53AZ B) SU03PS 12 3.74 1.50 3.33 0.89 7.081.64 0.89 4.7 0.1 670 95 10 64 26 FA03PS 37 4.14 1.64 5.57 0.70 9.711.76 1.35 4.7 0.2 779 74 12 63 20 Average/low pH, citric+ 52 4.24 3.878.11 4.8 701 11 67

TABLE 13B Hybrids of MEHARI, low pH/high pH Tot. tot. Hex/suc Growing nrsuc Suc hex hex Sugar sugar Inv pH pH citric citric citric/ malic malicconditions frt avg Stdev avg stdev Avg stdev activity avg stdev avgstdev malic avg stdev F1 Hybrid SP03PS 7 5.06 1.26 3.15 0.69 8.21 0.626.6 0.2 233 70 3 73 10 MEHARI FA03PS 7 2.86 0.80 6.53 1.42 9.40 1.702.28 6.0 0.2 316 137 9 34 23 High pH Average 14 3.96 4.84 8.80 1.22 6.3274 5 54 F1 Hybrid SP03PS 7 3.10 1.42 4.46 0.53 7.55 1.44 5.4 0.2 447 856 79 22 low pH, citric− FA03PS 6 1.39 1.46 5.75 0.66 7.14 1.99 4.14 4.60.2 527 80 7 76 27 Average 13 2.24 5.10 7.34 2.27 5.0 487 6 78 F1 HybridSP03PS 7 3.19 1.22 4.56 0.67 7.76 1.43 4.8 0.1 619 78 12 53 23 Low pH,citric+ SP03OF1 3 5.82 1.55 3.63 0.76 9.45 0.86 0.62 4.9 0.2 718 57 2037 7 FA03PS 7 1.12 0.52 6.73 0.56 7.84 0.88 6.02 4.5 0.1 654 128 7 92 69Average 17 3.37 4.97 8.35 1.47 4.76 663 11 60

Example 12

Sensory Analysis

Sensory analysis of fruits of plants of the instant invention wasperformed by an Expert panel. The panel was composed of 12 persons,especially trained to describe melons texture, sugar and acid savours.Six sessions of training were carried out before the expert sessions.The training program of the study was:

-   -   Training 1: Reading of the lexicon of tasting. Notation of 6        products in the assembly room and discussion in order to see        which descriptor is difficult to agree about.    -   Training 2: Notation of 2 products+2 repeated in laboratory of        sensory analysis to see how the panel evaluate the descriptors        personnally.    -   Training 3: Results of the training 2 are presented to the        panel. Notation of 2 products+2 repeated in the assembly room        and discussion. Notation of 2 products in the individual cabins.    -   Training 4: Works on sugar and acid solutions at different        concentrations to evaluate the difficulties of the panel on        these descriptors. Results of the training 3 are presented to        the panel. Notation of 3 products+1 repeated in the assembly        room and discussion. Notation of 3 products+1 repeated in        individual cabins.    -   Training 5: Notation of 4 products in the assembly room and        notation of 2 products+2 repeated in the individual cabins. It        allows us to check the repeatability of the tasters inside the        session. Works on acid solutions.    -   Training 6: Notation of 3 products in the assembly room and        notation of 2 products+2 repeated in the individual cabins to        check the repeatability of the tasters inside the session.

The sensory analysis was carried out in an air-conditioned sensoryanalysis laboratory equipped with individual cabins. A structured scaleof quotation in 10 points (from 0 to 9) was used with the followingdescriptors:

Sweet Savor

Definition: It is about the perception of the sweet savour perceived inmouth. Mode of evaluation: Chew the product until its disappearance.Estimate the intensity of the sweet savour. Notation: 0=not sweet,9=very sweet

Acid Savor

Definition: It is about the perception of the acid savour perceived inmouth. Mode of evaluation: Chew the product until its disappearance.Estimate the intensity of the acid savour. Notation: 0=not acid, 9=veryacid.

Protocol of Preparation

-   -   -   Wash of melons in the cold water        -   Longitudinal cutting of the melon        -   Removing of the pulp and the seeds        -   Cutting each half of melons in 3 parts (longitudinal            cutting)        -   Removing of the extremities of 3 parts        -   Cutting each part in 2 in order to have 6 parts on the half            of the melon            Presentation of the Products        -   Samples are presented coded with random numbers and            according to a plan of presentation (Latin Square) avoiding            the effects of presentation order.        -   The products are given one after the other.

Tables 14 and 15 show the results of two sensory analysis conducted ondifferent fruits. In Table 14, plants were sown in early March andtransplanted to open protected fields as vine crops in early April.Fruits were harvested in early July. In Table 14 A, fruits were storedfor 7 to 12 days at 10° C. followed by 2 days at 20° C. In Table 14 B,fruits were stored for 4 to 8 days at 10° C. followed by 2 days at 20°C.

In Table 15, plants were sown in early August and transplanted toplastic houses as staked crops in late August. Fruits were harvested inearly November. Fruits were stored for 4 to 8 days 5° C. followed by 3days at 20° C.

Sensory analysis was performed on the number of fruits indicated in theTables. Each fruit was tested by the 12 qualified tasters. The figuresfor acid and sugar savors for each line represent the average of thenumber of fruit times 12 (for the 12 tasters). Tables 14 and 15 alsoshow measurements of the pH and contents of sugars and organic acid forthe fruits tested. TABLE 14A FRT GLU SUC FRU Tot sug Citric Ac Malic AcACID SUGAR nr Brix pH g/100 g g/100 g g/100 g/100 g mg/100 g mg/100 gsavor savor YUSOL/1 X Avg 10 11.7 6.24 1.2 7.2 2.1 10.6 309 4 0.5 5.6SOLAZ/1 basic Std Dev 2.1 0.30 0.3 2.3 0.3 1.9 121 10 0.3 1.0 YUSAZA Avg6 12.7 4.79 1.1 7.7 2.0 10.7 808 0 2.1 5.4 X YUSOL Std Dev 0.5 0.09 0.20.6 0.2 0.5 17 0 0.7 0.5 SOLAZ/2 Avg 10 13.5 4.68 1.5 7.9 2.0 11.3 83218 3.6 4.7 X YUSOL/3 Std Dev 1.4 0.17 0.5 1.0 0.3 1.0 37 20 0.5 0.3SOLAZ/1 Avg 10 13.2 4.58 1.8 7.2 2.4 11.4 816 41 3.1 4.7 X YUSOL/1 StdDev 0.8 0.22 0.6 1.2 0.4 0.6 37 32 1.2 0.6 Average SOLAZ XYUSOL 20 13.44.63 1.7 7.6 2.2 11.4 824 30 3.4 4.7

TABLE 14B FRT GLU SUC FRU SUG Citric Ac Malic ACID SUGAR nr Brix pHg/100 g g/100 g g/100 g Tot mg/100 g mg/100 g savor savor Female MehariAvg 6 13.2 6.64 1.0 8.5 1.7 11.2 272 63 0.5 5.8 X L53 Std Dev 1.7 0.090.1 1.5 0.2 1.5 54 32 0.2 0.6 Female Mehari Avg 7 13.4 5.20 1.1 7.8 1.910.8 652 88 1.7 5.2 X L53 AZ A/1 Std Dev 0.7 0.11 0.2 0.8 0.3 0.9 87 690.4 0.4 Female Mehari Avg 5 13.1 5.28 1.2 8.1 2.0 11.3 581 107 1.6 5.3 XL53 AZ A/2 Std Dev 0.8 0.08 0.2 0.9 0.1 0.8 50 40 0.4 0.6 AVG FemaleMehari X L53 AZ A 12 13.3 5.24 1.2 8.0 2.0 11.1 617 98 1.7 5.3 FemaleMehari Avg 6 12.2 4.98 1.2 7.1 1.9 10.2 708 107 3.2 4.6 X L53 AZ B StdDev 1.0 0.21 0.1 0.7 0.2 0.9 78 44 0.6 0.3

TABLE 15 FRT GLU SUC FRU Tot sug Citric Ac Malic Ac ACID SUGAR nr BrixpH g/100g g/100 g g/100 g g/100g mg/100 g mg/100 g savor savor SOLAZ/2Avg 3 9.60 4.53 1.97 2.98 2.47 7.41 717 0 3.78 4.31 X YUSOL/3 Std Dev0.53 0.06 0.36 0.30 0.36 0.94 39 0 0.84 0.55 SOLAZ/1 Avg 2 11.00 4.742.14 4.04 3.05 9.24 574 0 2.54 5.63 X YUSOL/1 Std Dev 0.85 0.09 0.350.00 0.09 0.44 169 0 1.20 0.06 Average SOLAZ X YUSOL 5 10.2 4.6 2.0 3.42.7 8.1 660 0 3.3 4.8 YUSOLX Avg 5 12.40 4.71 3.04 4.02 3.59 10.65 71318 2.95 5.47 YUSAZA Std Dev 1.04 0.17 0.56 1.04 0.67 0.68 42 15 0.400.75 TD X L53 Avg 8 12.13 4.97 1.23 5.53 2.06 8.82 696 74 2.56 5.72 AZ/AStd Dev 1.13 0.14 0.36 1.13 0.32 0.80 83 41 0.85 0.39 Mehari female Avg3 9.50 5.10 1.48 3.48 2.52 7.48 632 62 2.17 5.50 X L53 AZ/A Std Dev 1.320.27 0.17 0.70 0.37 1.17 79 25 0.43 0.55

Example 13

Molecular Marker Analysis

DNA was extracted from young leaves (15 day-old seedlings). Leaves werefreeze-dried and DNA was extracted following the method of Dellaporta(Dellaporta 1983).

PCR cycling conditions were: 15 s denaturation at 94° C. followed by 15s annealing at 54° C. and 30 s extension at 72° C. for 40 cycles.Sample's DNA was initially denatured for 2 minutes at 94° C. andextended for 2 min at 72° C. after PCR.

The PCR mix contained 1.65 mM MgCl₂, 60 mM of each deoxyribonucleotide,1×Taq Buffer, 0.2 unit Taq polymerase, 15-20 ng template DNA and 400 nMof each non-fluorescent primer or 200 nM of each fluorescently labeledprimer. Fluorescent primers were labeled with 6-FAM, NED or HEX.Fluorescent PCR products were separated on an ABI3700 capillarysequencer and their sizes measured using Applied Biosystem's Genescanand Genotyper fragment analysis software.

Non-fluorescent PCR products were separated by electrophoresis in 3%agarose gels (Resophor, Eurobio) at 400V with cooling system. Gels werestained with ethidium bromide. Three markers were used, CMAT141(described in Danin-Poleg et al. (2001) Theor. Appl. Genet. 102: 61-72and Danin-Poleg et al. (2002) Euphytica 125: 373-384) and NE0585 andNE1746. The primers for these markers are shown below. Forward PrimerReverse Primer (5′-3′) (5′-3′) CMAT141 AAGCACACCACCACCCGTAAGTGAATGGTATGTTATCCTTG (SEQ ID NO:1) (SEQ ID NO:2) NE0585GTATCATGTCGGAGAAACG CCTTTATCCCCACTTTTTC (SEQ ID NO:3) (SEQ ID NO:4)NE1746 TTCTCCGATGTGTCCTCTC GTCGCTTGGAATATATCGG (SEQ ID NO:5) (SEQ IDNO:6)

Example 14

Analysis of Accessions

A number of melon accessions comprising a low pH trait were analyzedusing the markers above. The sizes of the amplified fragments arereported in Table 16 below. The indicated sizes (in bp) are not absolutebut relative to the other size products detected with the same primerpair. The real (exact) size of the amplified fragments (e.g. determinedby sequencing) could be slightly different (+/−1 bp) of those indicatedherein. TABLE 16 CMAT141 NE0585 NE1746 IND35-1.2. 173 230 127 IND35-2.3.168 232 124 FAGOUS2:4- 176 228 253 121 FAGOUS2:5- 176 121 142 FAGOUS2:7-176 253 142 PI414723 175 228 124 PI414724 175 124 PI161375 175 228 124PI124112 175 239 124

Example 15

Evolution of Fruit Characteristics of Fruits Kept on the Plant

Plants were grown as for the plants in Table 15 above. Fruits wereharvested at different maturation times points (cycle) calculated by thedifference between the harvest date (HDT) and the pollination date(PDT). Maturity was considered as reached at a particular time point inthe cycle, when the average of the sucrose content for fruits at theparticular time point in the cycle reached 2.0 g sucrose per 100 g fwtand using other ripening signs such as side leaf senescence. The resultsare shown in Table 17.

The results are based on individual fruit measurements and it isunderstood that some fruits may yield deviating values because ofdamages, aberrant development or other environmental factors. TABLE 17Flesh Orange FRT Colour GLU SUC FRU HEX Tot sug Citric Ac Malic Ac nrPDT HDT Cycle 1 to 5 pH Brix g/100 g g/100 g g/100 g g/100 g g/100 gmg/100 g mg/100 g TD X 1 17 37 20 1 5.09 5.0 1.6 0.0 1.4 2.6 3.0 169 460L53 AZ A/1 1 16 37 21 1 4.83 5.4 1.8 0.0 1.7 3.9 3.5 167 273 Maturity 114 37 23 1 4.94 4.6 1.4 0.0 1.4 2.7 2.8 128 243 reached between 1 16 4024 1 4.55 4.8 1.4 0.0 1.2 2.8 2.6 387 314 cycle 33 and 40 1 16 40 24 14.66 5.0 1.2 0.0 1.7 2.9 2.9 387 114 1 14 40 26 2 4.88 6.5 1.9 0.0 1.42.3 3.3 498 255 1 14 40 26 2 4.44 7.0 1.4 0.6 2.7 4.1 4.7 852 119 1 1444 30 2 4.36 6.7 2.3 0.4 2.0 3.2 4.6 776 178 1 14 44 30 2 4.47 7.0 1.70.5 2.2 3.9 4.3 528 77 1 14 44 30 2 4.70 7.0 2.3 0.2 2.4 4.7 5.0 649 111 14 47 33 2 4.37 6.5 2.0 0.0 1.6 3.2 3.6 730 174 1 14 47 33 2 4.59 7.01.2 1.0 1.7 2.9 3.9 453 117 1 14 47 33 3 4.85 13.6 2.3 3.9 1.9 4.0 8.2876 199 1 14 54 40 3 5.02 13.0 1.0 7.3 2.2 3.2 10.5 663 89 1 17 57 40 34.83 11.5 1.5 4.5 2.4 3.9 8.4 590 155 1 16 57 41 3 4.79 14.0 1.8 6.9 2.13.9 10.9 890 31 1 16 58 42 4 4.75 13.3 2.1 5.2 2.0 3.1 9.2 985 104 1 1457 43 3 4.96 12.7 1.7 5.5 1.9 4.3 9.0 893 167 1 14 57 43 4 5.24 14.0 1.48.4 2.3 3.7 12.1 648 40 1 14 57 43 4 5.06 12.5 1.0 5.9 1.8 2.8 8.7 62348 1 14 57 43 4 5.00 12.5 1.3 5.7 2.2 3.4 9.2 758 29 TD X 1 17 37 20 14.45 4.8 1.7 0.1 1.8 3.5 3.5 307 319 L53 AZ A/2 1 17 37 20 1 4.53 5.21.9 0.0 2.0 5.0 3.9 331 332 Maturity 1 14 37 23 1 4.43 4.1 1.0 0.0 1.12.3 2.1 572 323 reached between 1 17 40 23 3 4.55 7.0 1.3 0.8 1.7 3.03.8 682 120 cycle 34 and 38 1 14 40 26 2 4.48 6.0 1.3 0.3 1.8 3.2 3.5590 136 1 14 40 26 3 4.44 6.2 1.9 0.0 1.9 1.5 3.8 594 206 1 17 44 27 24.36 4.9 1.5 0.0 1.3 2.8 2.9 640 295 1 19 47 28 3 4.33 8.9 2.8 0.0 2.74.4 5.5 977 156 1 14 44 30 2 4.65 6.0 1.0 1.3 2.4 3.4 4.8 715 56 1 17 5134 3 4.43 10.1 2.9 1.0 2.9 5.9 6.8 989 141 1 17 51 34 3 4.77 11.0 1.93.5 2.5 4.4 7.9 836 31 1 17 51 34 3 4.91 10.5 1.3 5.3 1.9 3.2 8.5 753 961 16 51 35 3 4.31 8.6 2.2 0.9 2.1 5.0 5.3 970 216 1 16 54 38 3 4.76 14.12.3 5.5 2.2 3.8 10.1 983 164 1 14 54 40 4 4.80 14.8 2.1 7.0 2.1 4.0 11.3992 166 1 14 54 40 4 5.03 15.0 0.8 9.0 1.8 2.6 11.6 843 5 1 17 58 41 34.99 14.5 1.6 8.0 2.1 3.6 11.6 909 75 1 14 57 43 4 4.97 12.0 0.8 5.8 1.72.5 8.2 635 76 1 14 57 43 5 5.20 14.0 1.2 6.3 1.7 2.9 9.2 711 64 1 14 5844 4 5.06 16.0 1.6 8.5 1.7 3.4 11.9 986 50 FRT GLU SUC FRU HEX Tot sugCitric Ac Malic Ac nr PDT HDT Cycle Colour pH Brix g/100 g g/100 g g/100g g/100 g g/100 g mg/100 g mg/100 g Female Mehari 1 21 40 20 1 4.45 4.71.5 0.0 1.3 1.3 437 292 X L53 AZ A/1 1 17 37 20 1 4.62 4.5 1.1 0.1 1.42.5 2.6 295 98 Maturity 1 17 40 23 1 4.78 5.0 1.0 0.2 1.9 2.9 3.1 482 71reached between 1 14 40 26 2 4.54 5.0 1.1 0.1 1.6 2.7 2.8 397 81 cycle36 and 38 1 17 44 27 2 4.46 5.0 1.2 0.4 1.9 3.1 3.4 547 38 1 17 44 27 24.43 4.9 1.6 0.0 1.4 1.4 3.0 561 187 1 17 44 27 2 4.52 6.0 1.5 0.7 1.93.4 4.1 505 18 1 19 51 32 4 4.81 8.0 1.3 2.7 2.2 3.5 6.2 575 76 1 19 5132 2 4.50 6.2 2.0 0.0 1.6 2.8 3.6 592 75 1 19 51 32 3 4.76 7.0 1.5 1.42.0 3.5 4.9 465 105 1 14 50 36 3 5.99 8.6 1.5 2.5 1.6 2.6 5.6 414 259 116 53 37 5 6.28 9.0 1.4 3.0 1.8 3.3 6.3 297 166 1 17 54 37 4 5.57 8.51.7 2.6 2.6 4.3 6.9 309 261 1 17 54 37 2 5.68 8.4 1.6 1.7 1.4 2.6 4.8339 276 1 14 51 37 3 4.86 5.8 1.3 0.4 1.4 3.9 3.1 446 339 1 14 51 37 46.36 9.0 1.5 2.7 1.9 3.4 6.1 276 167 1 17 54 37 4 5.21 12.0 1.6 4.5 1.93.5 8.0 532 56 1 16 54 38 5 5.79 9.5 1.4 3.8 2.7 4.1 7.9 351 264 1 17 5740 2 5.47 9.0 1.7 1.5 1.4 4.0 4.6 410 312 1 17 57 40 4 4.63 10.9 2.4 3.02.3 3.8 7.7 894 111 1 17 57 40 5 6.22 9.5 1.5 3.8 2.1 3.6 7.4 360 277Female Mehari 1 17 37 20 1 4.84 4.5 0.7 0.3 1.7 2.5 2.7 368 33 X L53 AZA/2 1 17 37 20 1 5.25 4.7 1.6 0.0 1.5 2.5 3.1 210 375 Maturity 1 17 3720 1 5.31 4.3 1.4 0.0 1.2 2.6 2.6 200 288 reached between 1 17 44 27 24.52 4.9 1.7 0.0 1.4 2.4 3.1 320 190 cycle 35 and 37 1 17 44 27 2 4.495.0 0.8 0.3 1.7 2.5 2.8 423 5 1 17 44 27 2 6.69 5.3 1.5 0.0 0.8 3.4 2.4238 160 1 17 44 27 2 4.47 6.0 2.1 0.0 1.7 7.5 3.8 437 183 1 21 51 30 34.63 9.5 2.6 1.4 2.4 4.3 6.4 548 150 1 17 47 30 2 4.53 7.4 2.4 0.1 2.14.0 4.6 672 126 1 17 47 30 3 4.60 6.5 1.5 1.0 2.3 3.8 4.8 562 44 1 17 5134 2 4.68 6.5 2.0 0.2 1.9 2.6 4.1 713 99 1 17 51 34 3 4.50 9.9 2.7 1.52.5 4.9 6.7 944 118 1 16 51 35 4 5.35 10.5 1.7 4.0 2.9 4.6 8.6 723 32 116 51 35 4 4.77 11.7 2.2 3.2 2.2 3.7 7.6 904 133 1 14 50 36 2 5.84 7.11.4 0.6 1.5 2.6 3.5 224 168 1 14 50 36 4 6.09 7.0 1.1 2.1 2.1 3.2 5.3273 201 1 14 50 36 2 5.95 8.3 1.6 1.7 1.8 3.7 5.1 279 235 1 14 50 36 56.51 9.5 1.7 2.9 2.7 4.4 7.3 336 202 1 14 51 37 4 5.13 10.0 1.5 3.8 2.43.9 7.7 599 77 1 19 57 38 3 5.60 13.1 2.3 4.1 2.2 3.3 8.6 613 150 1 1957 38 3 4.62 12.2 3.0 3.7 2.7 5.2 9.4 1011 63 FRT GLU SUC FRU HEX Totsug Citric Ac Malic Ac nr PDT HDT Cycle pH Brix g/100 g g/100 g g/100 gg/100 g g/100 g mg/100 g mg/100 g SOLAZ/2 x2 9 37 28 4.23 5.0 1.6 0.11.7 4.0 3.3 664 116 X YUSOL/3 x2 9 37 28 4.30 5.1 1.8 0.0 1.9 4.8 3.7496 123 Maturity x2 9 37 28 4.21 5.9 2.4 0.0 2.0 3.7 4.4 674 77 reachedbetween 1 9 37 28 4.53 6.0 2.3 1.0 2.6 4.8 5.9 675 102 cycle 38 and 41x2 9 37 28 4.19 5.3 1.6 0.1 1.8 3.9 3.5 703 123 x2 9 37 28 4.37 4.6 1.40.0 1.4 2.9 2.8 509 264 x2 10 40 30 4.18 5.5 2.0 0.0 1.9 3.2 3.9 545 94x2 9 40 31 4.21 6.4 2.1 0.0 2.1 3.5 4.1 795 71 1x 9 40 31 4.25 7.2 2.90.0 2.9 4.8 5.8 723 186 1 9 40 31 4.37 6.8 2.3 0.3 2.8 5.1 5.4 667 55 x29 40 31 4.15 5.5 1.9 0.0 2.0 4.7 3.9 728 146 1x 9 44 35 4.21 8.6 2.8 0.03.3 6.4 6.2 990 34 x2 9 44 35 4.70 7.0 2.1 1.7 3.0 5.0 6.7 706 0 x2 9 4435 4.18 7.3 2.3 0.0 2.8 5.3 5.1 984 32 x2 9 44 35 4.17 7.3 2.1 0.0 2.75.8 4.8 992 28 x2 9 44 35 4.06 6.5 2.3 0.0 2.3 4.9 4.6 990 67 1 9 44 354.34 7.2 1.9 0.5 2.3 4.2 4.7 736 14 1 9 47 38 4.49 10.2 2.4 3.3 2.8 5.28.5 761 0 x2 9 47 38 4.24 9.0 2.6 0.6 3.1 6.7 6.2 782 15 x2 9 47 38 4.2110.5 3.1 0.9 3.4 7.3 7.5 834 41 1 9 47 38 4.60 9.2 1.7 2.7 2.5 4.2 6.9695 0 x2 9 47 38 4.21 10.3 2.9 1.2 3.0 6.2 7.1 1052 53 x2 9 50 41 4.449.2 2.2 2.0 2.4 4.7 6.6 701 24 x2 9 50 41 4.69 10.5 803 66 x2 9 50 414.76 10.2 853 37 x2 9 50 41 4.77 10.4 1.8 3.6 2.8 4.6 8.2 637 1 x2 9 5041 4.56 12.2 943 0 1 9 50 41 4.51 9.4 1.8 2.9 2.1 3.9 6.9 694 0 x2 9 5142 4.55 10.5 887 0 x2 9 54 45 4.28 12.4 2.7 3.3 2.6 5.0 8.7 982 48 x2 954 45 4.23 10.8 2.4 3.0 2.6 4.7 8.0 1026 61 1 9 54 45 4.44 12.9 2.7 4.72.4 3.6 9.8 997 52 SOLAZ/1 x2 10 37 27 4.36 4.8 1.7 0.0 1.7 3.7 3.3 526136 X YUSOL/1 1 10 37 27 4.84 4.0 1.4 0.4 2.1 3.5 3.9 428 27 Maturity 19 37 28 4.51 4.5 2.1 0.0 2.1 4.2 4.2 476 86 reached between 1 9 37 284.49 5.4 2.1 1.1 2.4 4.5 5.6 524 55 cycle 38 and 41 1 9 40 31 4.42 6.21.8 0.4 2.8 4.6 5.1 598 62 1 9 40 31 4.38 5.0 1.6 0.1 2.0 3.6 3.7 546 941 9 40 31 4.25 5.8 1.9 0.0 2.1 3.6 3.9 642 91 1 9 44 35 4.53 7.0 2.2 1.63.2 5.4 7.0 654 0 1 9 44 35 4.24 8.7 2.7 0.5 3.0 5.3 6.1 977 70 1 9 4435 4.44 7.6 2.0 0.8 2.6 4.6 5.4 498 0 1 9 44 35 4.38 8.6 2.3 1.4 3.1 5.56.9 640 6 1 9 47 38 4.42 10.4 1.8 2.4 2.6 4.4 6.9 733 0 1 9 47 38 4.538.0 2.0 1.8 3.0 4.9 6.7 540 0 1 9 47 38 4.74 8.5 2.2 1.6 2.7 5.4 6.5 6003 1 9 47 38 4.50 10.1 2.3 1.8 2.5 3.4 6.7 568 15 1 9 50 41 4.45 10.2 1.72.6 2.2 4.0 6.6 676 8 1 9 50 41 4.80 10.4 1.9 4.0 3.0 4.9 8.9 455 0 1 950 41 4.67 11.6 2.4 4.0 3.1 5.5 9.5 694 0 x2 9 51 42 4.42 11.1 796 28 19 51 42 4.75 8.6 767 8 1 9 54 45 4.40 11.2 2.2 2.5 2.4 4.4 7.1 745 19 19 54 45 4.47 8.5 1.8 1.3 1.9 3.8 5.1 705 11 YUSAZ A/1 1 14 37 25 5.227.1 2.5 0.0 2.6 5.1 5.1 284 406 X YUSOL/1 x2 10 37 27 4.38 5.9 2.1 0.01.8 6.7 3.9 513 146 Maturity x2 9 37 28 4.67 5.5 2.0 0.0 2.7 4.7 4.7 53093 reached between x2 9 37 28 4.37 6.2 2.1 0.0 2.0 4.0 4.1 695 104 cycle35 and 42 1 19 47 28 5.23 5.8 1.9 0.6 3.1 5.0 5.6 410 0 1 9 37 28 4.657.2 2.8 0.0 3.4 6.2 6.2 594 22 x2 9 40 31 4.66 7.0 2.0 0.4 2.8 4.8 5.2490 12 1 9 40 31 4.50 7.4 2.3 0.1 2.8 5.1 5.2 635 36 x2 9 40 31 4.32 5.92.2 0.0 2.1 4.0 4.3 495 102 x2 9 44 35 4.39 7.3 2.7 0.3 2.4 4.2 5.4 57955 1 9 44 35 4.62 9.2 2.5 1.2 3.0 5.5 6.7 689 0 1 9 44 35 4.39 8.4 3.00.0 3.5 6.7 6.6 685 55 x2 9 44 35 4.41 8.8 3.0 0.3 3.0 5.4 6.3 702 27 19 44 35 4.77 9.6 2.7 1.6 3.4 6.2 7.8 736 4 x2 9 44 35 5.85 8.0 1.9 3.13.4 5.3 8.4 387 0 x2 9 47 38 4.46 8.9 3.1 0.3 3.6 6.9 7.1 653 37 x2 9 4738 4.85 11.4 3.9 2.4 4.7 8.6 11.0 776 17 1 9 47 38 6.46 9.4 1.3 3.6 2.84.1 7.8 274 0 x2 9 47 38 4.75 9.2 2.8 1.4 3.1 5.9 7.3 525 0 1 9 51 424.87 12.4 2.8 4.3 3.1 5.9 10.2 696 19 x2 9 51 42 4.45 11.4 2.6 3.9 3.25.8 9.7 678 11 x2 9 51 42 4.57 14.4 791 47 x2 9 51 42 4.41 11.6 2.7 3.02.6 5.1 8.3 730 52 x2 9 51 42 4.73 13.8 2.5 5.3 3.5 6.0 11.3 679 0 1 1053 43 6.24 11.0 410 47 x2 10 53 43 6.18 10.5 350 48 1 9 54 45 4.58 13.52.7 4.8 2.8 4.8 10.4 780 62 1x 9 54 45 4.54 13.2 2.5 4.2 2.3 4.4 9.0 75134 YUSAZ A/2 x2 10 37 27 4.55 7.0 2.7 0.0 3.1 5.7 5.8 543 24 X YUSOL/3x2 10 37 27 4.52 7.6 2.8 0.0 3.1 6.0 5.9 640 64 Maturity 1 9 37 28 5.468.3 2.2 1.1 2.9 4.9 6.2 457 166 reached between x2 9 37 28 4.40 6.1 2.50.0 2.5 4.4 5.0 630 64 cycle 34 to 38 1 9 37 28 4.67 9.4 2.9 2.5 4.1 7.09.5 618 30 1 10 40 30 4.71 9.5 3.1 1.3 3.3 6.4 7.7 610 54 x2 9 40 314.51 6.6 2.4 0.0 2.7 5.2 5.1 460 64 x2 9 40 31 4.62 7.7 3.0 0.0 3.2 6.16.1 569 50 x2 9 40 31 4.58 7.6 2.7 0.0 3.2 5.2 6.0 655 41 x2 9 40 314.48 8.3 2.9 0.0 3.4 2.6 6.3 721 48 1 10 44 34 4.76 11.2 3.4 1.0 4.0 7.28.5 844 74 1 10 44 34 4.69 13.1 3.1 3.1 3.9 9.7 10.0 825 90 1 17 51 344.72 10.1 612 22 x2 10 44 34 4.48 12.1 3.4 2.0 4.0 6.7 9.4 683 64 1 9 4435 4.66 13.0 3.3 4.2 3.5 6.8 11.0 733 42 1 9 47 38 5.22 14.4 2.6 5.9 3.05.6 11.4 514 87 1 9 47 38 4.50 13.2 3.1 4.1 3.1 5.8 10.3 899 80 x2 9 4738 4.89 12.4 2.2 4.1 3.2 6.7 9.5 696 21 1 9 47 38 4.57 14.2 3.3 4.3 3.36.2 10.9 725 66 1 9 47 38 4.66 15.2 2.8 5.7 2.9 4.8 11.4 774 141 x2 1051 41 4.67 15.2 818 44 x2 9 51 42 4.74 13.6 754 65 1 9 51 42 4.97 15.42.1 7.0 2.6 4.7 11.7 507 1 1 9 51 42 5.60 15.2 2.0 8.6 2.3 4.3 12.9 46297 1 11 54 43 4.94 15.8 2.9 6.0 2.8 8.1 11.7 680 66 x2 9 53 44 4.93 15.1881 221 x2 9 54 45 4.80 15.3 2.3 7.4 2.7 5.1 12.4 831 103 x2 9 54 454.53 14.1 2.7 5.2 2.8 5.2 10.8 876 64 1 9 54 45 5.00 15.8 2.8 6.3 2.54.8 11.6 641 188

Example 16

Post Harvest Evolution of Fruit Characteristics

A. Fruits of plants of trial SP03OF (see Tables 6 and 7C) were harvestedand stored for 1 week at 12° C. followed by 3 days at 20° C. or for 6weeks at 12° C. followed by 2 days at 20° C. The characteristics of thefruits were measured after storage and are shown in Table 18.

B. Plants were grown as for the plants in Table 14 above. Fruits wereharvested and stored for 1 week at 12° C. followed by 3 days at 20° C.The characteristics of the fruits were measured after storage and areshown in Table 19. TABLE 18 FRT Glu Suc Fru Hex Total Citric Malic Postharvest protocol Hv Nr Brix pH g/100 g g/100 g g/100 g g/100 g Sugarmg/100 g mg/100 g Millenium 1 w 12° + 3 d 20° Avg 5 8.4 5.75 2.0 2.3 2.24.2 6.4 441 −8 Stdev 0.3 0.05 0.1 0.2 0.0 0.1 0.2 25 4 6 w 12° + 2 d 20°Avg 2 10.1 5.86 1.5 3.9 2.2 3.7 7.6 487 Stdev 0.6 0.03 0.3 0.9 0.2 0.50.4 4 YUSOL/1&2 1 w 12° + 3 d 20° Avg 4 12.0 4.71 1.0 6.7 2.1 3.1 9.8802 −1 X SOLAZ/1 Stdev 0.7 0.05 0.1 0.7 0.1 0.2 0.7 33 6 6 w 12° + 2 d20° Avg 2 13.2 4.77 0.5 8.1 1.8 2.3 10.4 808 Stdev 0.5 0.07 0.1 0.6 0.00.1 0.5 36 YUSOL/3 6 w 12° + 2 d 20° Avg 4 11.9 4.89 1.1 6.2 1.9 3.0 9.2638 X SOLAZ/1 Stdev 0.9 0.07 0.4 1.6 0.3 0.7 1.1 72 YUSOL/1 1 w 12° + 3d 20° Avg 9 9.5 4.46 1.0 4.5 1.9 2.9 7.4 773 −5 X SOLAZ/2 Stdev 1.4 0.120.2 1.5 0.2 0.4 1.2 85 5 6 w 12° + 2 d 20° Avg 4 9.5 4.65 1.1 3.7 2.03.1 6.7 669 Stdev 1.4 0.17 0.2 1.4 0.3 0.5 1.0 70 YUSOL/2 1 w 12° + 3 d20° Avg 5 10.1 4.53 1.1 4.9 2.1 3.2 8.2 789 3 X SOLAZ/2 Stdev 2.3 0.130.2 2.3 0.2 0.3 2.1 95 6 w 12° + 2 d 20° Avg 2 10.6 4.70 1.1 5.5 2.0 3.18.6 697 Stdev 0.7 0.02 0.1 0.6 0.1 0.2 0.4 51 YUSOL/3 1 w 12° + 3 d 20°Avg 12 10.4 4.50 0.9 5.8 1.8 2.7 8.4 777 −4 X SOLAZ/2 Stdev 0.8 0.09 0.20.9 0.1 0.3 0.8 37 6 6 w 12° + 2 d 20° Avg 6 11.0 4.85 0.6 6.2 1.5 2.18.3 656 Stdev 0.8 0.14 0.3 1.1 0.3 0.6 0.7 46

TABLE 19 ratio FRT GLU SUC FRU Tot sug Citric Ac Malic Ac Citric Ac/ NrBrix pH g/100 g g/100g g/100 g g/100 g mg/100 g mg/100 g Malic AcSOLAZ/2 X YUSOL/3 51 13.1 4.75 1.4 7.2 2.0 10.6 771 23 35 SOLAZ/1 XYUSOL/1 51 13.8 4.71 1.8 7.9 2.3 12.0 778 21 53 YUSOL/1 X 51 13.4 6.141.4 8.0 2.3 11.7 354 14 31 SOLAZ/1 basic MEHARY 10 14.8 6.51 0.5 10.41.3 12.2 289 73 4 Female MEHARI 59 12.4 5.45 0.8 7.4 1.4 9.7 491 211 2 XL53 AZ/A TD x L53AZ/A 49 13.9 5.16 0.6 9.0 1.3 10.9 579 69 11

1. A C. melo plant capable of producing fruit, wherein said fruitcomprises at maturity: a) at least about 400 mg citric acid per 100 gfwt; b) pH of about 4.2 to about 5.6; and c) at least about 5.0 g sugarper 100 g fwt.
 2. The C. melo plant according to claim 1, wherein theratio of citric acid to malic acid in said fruit is greater than 4.4. 3.The C. melo plant according to claim 1, wherein said fruit comprisesless than about 85 mg malic acid per 100 g fwt.
 4. The C. melo plantaccording to claim 1, wherein said fruit comprises an average acid savorof about 1.6 to about 3.8 and an average sugar savor of about 4.3 toabout 5.8.
 5. The C. melo plant according to claim 4, wherein said fruitcomprises an average acid savor of about 2.5 to about 3.8 and an averagesugar savor of about 4.3 to about 5.6.
 6. The C. melo plant according toclaim 4, wherein said fruit comprises an average acid savor of about 1.6to about 3.0 and an average sugar savor of about 5.2 to about 5.8. 7.The C. melo plant according to claim 1, wherein a DNA fragment of about168 bp to about 178 bp is amplified from DNA of said plant when theprimers of marker CMAT141 are used.
 8. The C. melo plant according toclaim 7, wherein said DNA fragment is 168 bp, 173 bp, 169 bp, 172 bp or178 bp long.
 9. The C. melo plant according to claim 1, wherein a DNAfragment of about 218 bp to about 253 bp is amplified from DNA of saidplant when the primers of marker NE0585 are used.
 10. The C. melo plantaccording to claim 9, wherein said DNA fragment is of 230 bp, 232 bp,218 bp, 229 bp, 234 bp or 239 bp long.
 11. The C. melo plant accordingto claim 1, wherein a DNA fragment of about 121 bp to about 145 bp isamplified from DNA of said plant when the primers of marker NE1746 areused.
 12. The C. melo plant according to claim 11, wherein said DNAfragment is 124 bp, 127 bp, 133 bp, 142 bp or 145 bp long.
 13. The C.melo plant according to claim 1, wherein the pH of said fruit remainswithin said range of about 4.2 to about 5.6 for at least 2 days aftersaid fruit reaches maturity when said fruit is kept on the plant. 14.The C. melo plant according to claim 1, wherein the pH of said fruitremains within said range of about 4.2 to about 5.6 for at least 3 daysafter said fruit reaches maturity when said fruit is kept on the plant.15. The C. melo plant according to claim 1, wherein the pH of said fruitremains within said range of about 4.2 to about 5.6 when said fruit isharvested and kept in storage for at least 5 days at 20° C. or for atleast 7 days at 8-12° C. followed by 3 days at 20° C.
 16. The C. meloplant according to claim 1, wherein the pH of said fruit remains withinsaid range of about 4.2 to about 5.6 when said fruit is harvested andkept in storage for at least 7 days at 20° C. or for at least 12 days at8-12° C. followed by 3 days at 20° C.
 17. The C. melo plant according toclaim 1, wherein the citric acid of said fruit remains at or above about400 mg per 100 g fwt for at least 2 days after said fruit reachesmaturity when said fruit is kept on the plant.
 18. The C. melo plantaccording to claim 1, wherein the citric acid of said fruit remains ator above about 400 mg per 100 g fwt for at least 3 days after said fruitreaches maturity when said fruit is kept on the plant.
 19. The C. meloplant according to claim 1, wherein the citric acid of said fruitremains at or above about 400 mg per 100 g fwt when said fruit isharvested and kept in storage for at least 5 days at 20° C. or for atleast 7 days at 8-12° C. followed by 3 days at 20° C.
 20. The C. meloplant according to claim 1, wherein the citric acid of said fruitremains at or above about 400 mg per 100 g fwt when said fruit isharvested and kept in storage for at least 7 days at 20° C. or for atleast 12 days at 8-12° C. followed by 3 days at 20° C.
 21. The C. meloplant according to claim 2, wherein the ratio of citric acid to malicacid of said fruit remains greater than 4.4 for at least 2 days aftersaid fruit reaches maturity when said fruit is kept on the plant. 22.The C. melo plant according to claim 2, wherein the ratio of citric acidto malic acid of said fruit remains greater than 4.4 for at least 3 daysafter said fruit reaches maturity when said fruit is kept on the plant.23. The C. melo plant according to claim 2, wherein the ratio of citricacid to malic acid of said fruit remains greater than 4.4 when saidfruit is harvested and kept in storage for at least 5 days at 20° C. orfor at least 7 days at 8-12° C. followed by 3 days at 20° C.
 24. The C.melo plant according to claim 2, wherein the ratio of citric acid tomalic acid of said fruit remains greater than 4.4 when said fruit isharvested and kept in storage for at least 7 days at 20° C. or for atleast 12 days at 8-12° C. followed by 3 days at 20° C.
 25. The C. meloplant according to claim 1, wherein said plant is capable of producing alow-turning or a non-turning fruit.
 26. The C. melo plant according toclaim 1, wherein said plant is capable of producing a low-climacteric ora non-climacteric fruit.
 27. The C. melo plant according to claim 1,wherein the flesh of said fruit is orange, white, green or yellow. 28.The C. melo plant according to claim 1, wherein the mesocarp of saidfruit represents more than 50% of the total fresh weight of said fruit.29. The C. melo plant according to claim 1, wherein said C. melo plantcomprises a low pH gene.
 30. The C. melo plant according to claim 1,wherein said low pH gene is homozygous or heterozygous in said plant.31. The C. melo plant according to claim 1, wherein said low pH trait isobtainable from a plant of line IND-35, representative seeds of which isdeposited under Accession number NCIMB 41202, or a descendent of saidline IND-35.
 32. The C. melo plant according to claim 1, wherein saidfruit comprises at maturity: a) about 600 mg to about 1,200 mg citricacid per 100 g fwt; b) pH of about 4.2 to about 5.1; c) about 5.0 g toabout 15.0 g sugar per 100 g fwt.
 33. The C. melo plant according toclaim 1, wherein said fruit comprises at maturity: a) about 400 mg toabout 650 mg citric acid per 100 g fwt; b) pH of about 4.6 to about 5.6;and c) about 5.0 g to about 15.0 g sugar per 100 g fwt.
 34. The C. meloplant according to claim 1, wherein said plant is an inbred line. 35.The C. melo plant according to claim 1, wherein said plant is a hybrid.36. The C. melo plant according to claim 1, wherein said plant is adihaploid.
 37. Seed of a C. melo plant according to claim
 1. 38. Fruitof a C. melo plant according to claim
 1. 39. Flesh of a C. melo plantaccording to claim
 1. 40. A method of increasing the citric acid contentof a fruit of a first C. melo plant comprising: a) obtaining a first C.melo plant; b) crossing said first C. melo plant with a second C. meloplant comprising a low pH trait; c) obtaining a progeny C. melo plant;d) determining the pH of a fruit of said progeny C. melo plant, and e)selecting a fruit of said progeny C. melo plant which has increasedcitric acid content and lower pH, when compared to a fruit of said firstC. melo plant.
 41. The method according to claim 40, wherein the citricacid content of a fruit of said progeny C. melo plant is increased byabout 1.5 to about 3 times and the pH of said fruit reduced by about 1to about 2.5 pH units, when compared to a fruit of said first C. meloplant.
 42. The method of claim 40, wherein said second C. melo plant isa plant of line IND-35, representative seeds of which is deposited underAccession number NCIMB 41202, or a descendent of said line IND-35.
 43. AC. melo plant capable of producing a fruit, wherein said fruit comprisesat maturity an average acid savor of about 1.6 to about 3.8 and anaverage sugar savor of about 4.3 to about 5.8.
 44. The C. melo plantaccording to claim 43, wherein said fruit comprises at maturity anaverage acid savor of about 2.5 to about 3.8 and an average sugar savorof about 4.3 to about 5.6.
 45. The C. melo plant according to claim 43,wherein said fruit comprises at maturity an average acid savor of about1.6 to about 3.0 and an average sugar savor of about 5.2 to about 5.8.46. The C. melo plant according to claim 43, wherein said fruit has a pHof about 4.2 to about 5.6.
 47. The plant according to claim 43, whereinsaid fruit has a citric acid content of at least about 400 mg per 100 gfwt.
 48. The plant according to claim 43, wherein said fruit has a sugarcontent of at least about 5.0 g sugar per 100 g fwt.
 49. Seed of a C.melo plant according to claim
 43. 50. Fruit of a C. melo plant accordingto claim
 43. 51. Flesh of a C. melo plant according to claim 43.